Salts of potassium atp channel openers and uses thereof

ABSTRACT

Provided are immediate or prolonged administration of certain salts of K ATP  channel openers such as diazoxide to a subject to achieve novel pharmacodynamic, pharmacokinetic, therapeutic, physiological, metabolic and compositional outcomes in the treatment of diseases or conditions involving K ATP  channels. Also provided are pharmaceutical formulations, methods of administration and dosing of the salts that achieve these outcomes and reduce the incidence of adverse effects in treated individuals. Further provided are method of co-administering the salts with other drugs to treat diseases of humans and animals.

RELATED PATENT APPLICATIONS

This application is a Continuation of U.S. application Ser. No.12/534,018, filed Jul. 31, 2009, which is a divisional application ofU.S. application Ser. No. 12/391,990 (now U.S. Pat. No. 7,572,789),filed Feb. 24, 2009, which is a Continuation of U.S. application Ser.No. 11/614,044 (now U.S. Pat. No. 7,799,777), filed Dec. 20, 2006, whichclaims the benefit of U.S. Provisional Application No. 60/854,740 filedOct. 27, 2006, and U.S. Provisional Application No. 60/756,941 filedJan. 5, 2006, all of which are incorporated herein by reference in theirentireties and for all purposes.

FIELD OF THE INVENTION

The present invention relates to salts of potassium ATP (K_(ATP))channel openers, methods of preparing such salts, and methods of usethereof for treatment of a variety of diseases and conditions, includingfor example, diabetes and obesity.

BACKGROUND OF THE INVENTION

The following description of the background of the invention is providedas an aid in understanding the invention and is not admitted to describeor constitute prior art to the invention.

ATP-sensitive potassium (K_(ATP)) channels play important roles in avariety of tissues by coupling cellular metabolism to electricalactivity. The K_(ATP) channel has been identified as an octamericcomplex of two unrelated proteins, which assemble in a 4:4stoichiometry. The first is a pore forming subunit, Kir6.x, which formsan inwardly rectifying K⁺ channel; the second is an ABC (ATP bindingcassette) transporter, also known as the sulfonylurea receptor (SURx)(Babenko et al., Annu. Rev. Physiol., 60:667-687 (1998)). The Kir6.xpore forming subunit is common for many types of K_(ATP) channels, andhas two putative transmembrane domains (identified as TM1 and TM2),which are linked by a pore loop (H5). The subunit that comprises the SURreceptor includes multiple membrane-spanning domains and twonucleotide-binding folds.

According to their tissue localization, K_(ATP) channels exist indifferent isoforms or subspecies resulting from the assembly of the SURand Kir subunits in multiple combinations. The combination of the SUR1with the Kir6.2 subunits (SUR1/Kir6.2) typically forms the adipocyte andpancreatic B-cell type K_(ATP) channels, whereas the SUR2A/Kir6.2 andthe SUR2B/Kir6.2 or Kir6.1 combinations typically form the cardiac typeand the smooth muscle type K_(ATP) channels, respectively (Babenko etal., Annu. Rev. Physiol., 60:667-687 (1998)). There is also evidencethat the channel may include Kir2.x subunits. This class of potassiumchannels are inhibited by intracellular ATP and activated byintracellular nucleoside diphosphates. Such K_(ATP) channels link themetabolic status of the cells to the plasma membrane potential and inthis way play a key role in regulating cellular activity. In mostexcitatory cells, K_(ATP) channels are closed under normal physiologicalconditions and open when the tissue is metabolically compromised (e.g.when the (ATP:ADP) ratio falls). This promotes K⁺ efflux and cellhyperpolarization, thereby preventing voltage-operated Ca²⁺ channels(VOCs) from opening. (Prog. Res Research, (2001) 31:77-80).

Potassium channel openers (PCOs or KCOs; also referred to as channelactivators or channel agonists), are a structurally diverse group ofcompounds with no apparent common pharmacophore linking their ability toantagonize the inhibition of K_(ATP) channels by intracellularnucleotides. Diazoxide is a PCO that stimulates K_(ATP) channels inpancreatic β-cells (see Trube et al., Pfluegers Arch Eur J Physiol, 407,493-99 (1986)). Pinacidil and chromakalim are PCOs that activatesarcolemmal potassium channels (see Escande et al., Biochem Biophys ResCommun, 154, 620-625 (1988); Babenko et al., J Biol Chem, 275(2),717-720 (2000)). Responsiveness to diazoxide has been shown to reside inthe 6^(th) through 11^(th) predicted transmembrane domains (TMD6-11) andthe first nucleotide-binding fold (NBF1) of the SUR1 subunit.

Diazoxide, which is a nondiuretic benzothiadiazine derivative having theformula 7-chloro-3-methyl-2H-1,2,4-benzothiadiazine 1.1-dioxide(empirical formula C₈H₇ClN₂O₂S), is commercialized in three distinctformulations to treat two different disease indications: (1)hypertensive emergencies and (2) hyperinsulinemic hypoglycemicconditions. Hypertensive emergencies are treated with Hyperstat IV, anaqueous formulation of diazoxide for intravenous use, adjusted to pH11.6 with sodium hydroxide. Hyperstat IV is administered as a bolus doseinto a peripheral vein to treat malignant hypertension or sulfonylureaoverdose. In this use, diazoxide acts to open potassium channels invascular smooth muscle, stabilizing the membrane potential at theresting level, resulting in vascular smooth muscle relaxation.

Hyperinsulinemic hypoglycemic conditions are treated with Proglycem®, anoral pharmaceutical version of diazoxide useful for administration toinfants, children and adults. It is available as a chocolate mintflavored oral suspension, which includes 7.25% alcohol, sorbitol,chocolate cream flavor, propylene glycol, magnesium aluminum silicate,carboxymethylcellulose sodium, mint flavor, sodium benzoate,methylparaben, hydrochloric acid to adjust the pH, poloxamer 188,propylparaben and water. Diazoxide is also available as a capsule with50 or 100 mg of diazoxide including lactose and magnesium stearate.

Several experimental formulations of diazoxide have been tested inhumans and animals. These include an oral solution tested inpharmacodynamic and pharmacokinetic studies and a tablet formulationunder development in the early 1960's as an anti-hypertensive, but nevercommercialized (see Calesnick et al., J. Pharm. Sci. 54:1277-1280(1965); Reddy et al., AAPS Pharm Sci Tech 4(4):1-98, 9 (2003); U.S. Pat.No. 6,361,795).

Current oral formulations of diazoxide are labeled for dosing two orthree times per day at 8 or 12 hour intervals. Most subjects receivingdiazoxide are dosed three times per day. Commercial and experimentalformulations of diazoxide are characterized by rapid drug releasefollowing ingestion with complete release in approximately 2 hours.Unless indicated differently, the term “approximately” when used in thecontext of a numeric value, refer to the stated numeric value +/−10%. Inthe context of two-theta angles from XRPD studies, the termapproximately refers to +/−5% of the stated numeric value.

Current oral formulations of diazoxide in therapeutic use result in arange of adverse side effects including dyspepsia, nausea, diarrhea,fluid retention, edema, reduced rates of excretion of sodium, chloride,and uric acid, hyperglycemia, vomiting, abdominal pain, ileus,tachycardia, palpitations, and headache. (See e.g., current packaginginsert for the Proglycem®). Oral treatment with diazoxide is used inindividuals experiencing serious disease where failure to treat resultsin significant morbidity and mortality. The adverse side effects fromoral administration are tolerated because the benefits of treatment aresubstantial. The adverse side effects profile of oral diazoxide limitthe utility of the drug in treating obese subjects at doses within thelabeled range of 3 to 8 mg/kg per day.

The effect of diazoxide in animal models of diabetes and obesity (e.g.obese and lean Zucker rats) has been previously reported. See e.g.Alemzadeh et al., Endocrinology 133:705-712 (1993); Alemzadeh et al.,Metabolism 45:334-341 (1996); Alemzadeh et al., Endocrinology140:3197-3202 (1999); Stanridge et al., FASEB J 14:455-460 (2000);Alemzadeh et al., Med Sci Monit 10(3): BR53-60 (2004); Alemzadeh et al.,Endocrinology 145(12):3476-3484 (2004); Aizawa et al., J of Pharma ExpTher 275(1): 194-199 (1995); and Surwit et al., Endocrinology141:3630-3637 (2000).

The effect of diazoxide in humans with obesity or diabetes has beenpreviously reported. See e.g., Wigand et al., Diabetes 28(4):287-291(1979), evaluation of diazoxide on insulin receptors; Ratzmann et al.,Int J Obesity 7(5):453-458 (1983), glucose tolerance and insulinsensitivity in moderately obese patients; Marugo et al., Boll Spec ItBiol Sper 53:1860-1866 (1977), moderate dose diazoxide treatment onweight loss in obese patients; Alemzadeh et al., J Clin Endocr Metab83:1911-1915 (1998), low dose diazoxide treatment on weight loss inobese hyperinsulinemic patients; Guldstrand et al., Diabetes andMetabolism 28:448-456 (2002), diazoxide in obese type II diabeticpatients; Ortqvist et al., Diabetes Care 27(9):2191-2197 (2004),beta-cell function measured by circulating C-peptide in children atclinical onset of type 1 diabetes; Bjork et al., Diabetes Care21(3):427-430 (1998), effect of diazoxide on residual insulin secretionin adult type I diabetes patients; and Qvigstad et al., DiabeticMedicine 21:73-76 (2004).

U.S. Pat. No. 5,284,845 describes a method for normalizing blood glucoseand insulin levels in an individual exhibiting normal fasting bloodglucose and insulin levels and exhibiting in an oral glucose tolerancetest, elevated glucose levels and at least one insulin level abnormalityselected from the group consisting of a delayed insulin peak, anexaggerated insulin peak and a secondary elevated insulin peak.According to this reference, the method includes administering diazoxidein an amount from about 0.4 to about 0.8 mg/kg body weight before eachmeal in an amount effective to normalize the blood glucose and insulinlevels.

U.S. Pat. No. 6,197,765 describes administration of diazoxide fortreatment for syndrome-X, and resulting complications, that includehyperlipidemia, hypertension, central obesity, hyperinsulinemia andimpaired glucose tolerance. According to this reference, diazoxideinterferes with pancreatic islet function by ablating endogenous insulinsecretion resulting in a state of insulin deficiency and high bloodglucose levels equivalent to that of diabetic patients that depend onexogenous insulin administration for normalization of their bloodglucose levels.

U.S. Pat. No. 2,986,573 describes the preparation of diazoxide and itsuse for the treatment of hypertension. The patent asserts that alkalimetal salts may be prepared by methods well-known in the art for thepreparation of a salt of a strong base with a weak acid. It also allegesa specific method for making a sodium salt of diazoxide. This patentdoes not provide any evidence to support the formation of any salt ofdiazoxide.

U.S. Pat. No. 5,629,045 describes diazoxide for topical ophthalmicadministration.

WO 98/10786 describes use of diazoxide in the treatment of X-syndromeincluding obesity associated therewith.

U.S. Patent publication no. 2003/0035106 describes diazoxide containingcompounds for reducing the consumption of fat-containing foods.

U.S. Patent Publication No. 2004/0204472 describes the use of a Cox-2inhibitor plus diazoxide in the treatment of obesity. Also describedtherein is the use of a Cox-2 inhibitor plus a pharmaceuticallyacceptable salt of diazoxide, wherein acceptable cations include alkalimetals and alkaline earth metals.

U.S. Patent Publication No. 2002/0035106 describes use of K_(ATP)channel agonists for reducing the consumption of fat containing food.This application mentions pharmaceutically acceptable acid additionsalts, pharmaceutically acceptable metal salts and optionally alkylatedammonium salts, but does not disclose or describe how to prepare anysuch salts. This patent also does not provide any evidence to supportthe formation of any salt of a K_(ATP) channel agonist.

U.K. Patent GB982072 describes the preparation and use of diazoxide andderivatives for the treatment of hypertension and peripheral vasculardisorders. This patent mentions non-toxic alkali metals salts but doesnot disclose or describe how to prepare any such salts. This patent doesnot provide any evidence to support the formation of any salt ofdiazoxide or its derivatives.

SUMMARY OF THE INVENTION

The current invention relates to methods of preparation and use ofalkali metal, tertiary amine and ammonium salts of diazoxide anddiazoxide derivatives. It has been surprisingly found that it isdifficult to produce salts of diazoxide and derivatives. In particular,the inventors have been unable to reproduce formation of a diazoxidesalt using the method asserted in U.S. Pat. No. 2,986,573. Contrary towhat is reported in the literature, salt formation with diazoxide andderivatives depends on a proper selection of solvent and counter-ion.

Provided herein are pharmaceutical formulations of K_(ATP) channelopeners and their use for treatment of various diseases and conditionsincluding diabetes and obesity. Such formulations are characterized asbeing bioavailable. A K_(ATP) channel opener as used herein has any oneor more of the following properties: (1) opening SURx/Kir6.y potassiumchannels, where x=1, 2A or 2B and y=1 or 2; (2) binding to the SURxsubunit of K_(ATP) channels; and (3) inhibiting glucose induced releaseof insulin following administration of the compound in vivo. Preferably,K_(ATP) channel openers are K_(ATP) channel openers with all threeproperties. K_(ATP) channel openers as defined herein are preferablysalts prepared from the compounds of Formulae I-VIII, as set forthbelow.

The present invention also provides salts of the compounds defined byFormulae I-VIII. Salts of Formulae I-IV provided herein includemonovalent alkali metal salts and monovalent and divalent salts oforganic compounds, preferably organic compounds which include anammonium moiety. Salts of Formulae V-VIII are also provided herein,preferably prepared with monovalent and divalent counter-ions.

K_(ATP) channel openers defined by Formula I are as follows:

wherein:

-   -   R¹ is selected from the group consisting of hydrogen, lower        alkyl, substituted lower alkyl, cycloalkyl, and substituted        cycloalkyl provided however that when R¹ is a substituted lower        alkyl or a substituted cycloalkyl, then the substituent does not        include an amino group;    -   R^(2a) is hydrogen;    -   X is a 1, 2 or 3 atom chain, wherein each atom is independently        selected from carbon, sulfur and nitrogen, and each atom is        optionally substituted with halogen, hydroxyl, lower alkyl,        substituted lower alkyl, lower alkoxy, cycloalkyl, substituted        cycloalkyl, or substituted lower alkoxy, provided however that        when an atom of the chain is substituted with substituted lower        alkyl, substituted lower alkoxy or substituted cycloalkyl, then        the substituent does not include an amino group;    -   wherein ring B is saturated, monounsaturated, polyunsaturated or        aromatic;    -   and all bioequivalents including salts, prodrugs and isomers        thereof.

In particular embodiments of Formula I, X is C(R^(a))C(R^(b)), whereinR^(a) and R^(b) are independently selected from the group consisting ofhydrogen, halogen, lower alkyl, substituted lower alkyl, cycloalkyl,substituted cycloalkyl, lower alkoxy, substituted lower alkoxy,sulfonyl, and the like. In further embodiments, R^(a) and R^(b) areindependently selected from the group consisting of hydroxyl,substituted oxy, substituted thiol, alkylthio, substituted alkylthio,sulfinyl, sulfonyl, substituted sulfinyl, substitutedsulfonylalkylsulfinyl, alkylsulfonyl, and the like. In a preferredembodiment, Ring B does not include any heteroatoms.

Salts of embodiments of the channel openers defined by Formula I may beprepared from the following: (a) metal hydroxides, preferably alkalimetal hydroxides (e.g., NaOH and KOH) and (b) organic hydroxides,preferably organic compounds which include at least one tertiary amineor at least one quaternary ammonium ion (e.g., diethylaminoethanol,triethylamine, hydroxyethylpyrrolidine, choline andhexamethylhexamethylenediammonium, and the like).

K_(ATP) channel openers defined by Formula II are as follows:

wherein:

-   -   R¹ is selected from the group consisting of hydrogen, lower        alkyl, substituted lower alkyl, cycloalkyl, and substituted        cycloalkyl provided however that when R¹ is a substituted lower        alkyl or a substituted cycloalkyl, then the substituent does not        include an amino group;    -   R^(2b) is hydrogen;    -   X is a 1, 2 or 3 atom chain, wherein each atom is independently        selected from carbon, sulfur and nitrogen, and each atom is        optionally substituted with halogen, hydroxyl, lower alkyl,        substituted lower alkyl, lower alkoxy, cycloalkyl, substituted        cycloalkyl, or substituted lower alkoxy, provided however that        when an atom of the chain is substituted with substituted lower        alkyl, substituted cycloalkyl or substituted lower alkoxy, then        the substituent does not include an amino group;    -   wherein ring B is saturated, monounsaturated, polyunsaturated or        aromatic;    -   and all bioequivalents including salts, prodrugs and isomers        thereof.

In particular embodiments of Formula II, X is C(R^(a))C(R^(b)), whereinR^(a) and R^(b) are independently selected from the group consisting ofhydrogen, halogen, lower alkyl, substituted lower alkyl, cycloalkyl,substituted cycloalkyl, lower alkoxy, substituted lower alkoxy,sulfonyl, and the like. In further embodiments, R^(a) and R^(b) areindependently selected from the group consisting of hydroxyl,substituted oxy, substituted thiol, alkylthio, substituted alkylthio,sulfinyl, sulfonyl, substituted sulfinyl, substituted sulfonyl,alkylsulfinyl, alkylsulfonyl, nitro and the like. In preferredembodiment, Ring B does not include any heteroatoms.

Salts of embodiments of the channel openers defined by Formula II may beprepared from the following: (a) metal hydroxides, preferably alkalimetal hydroxides (e.g., NaOH and KOH) and (b) organic hydroxides,preferably organic compounds which include at least one tertiary amineor at least one quaternary ammonium ion (e.g., diethylaminoethanol,triethylamine, hydroxyethylpyrrolidine, choline andhexamethylhexamethylenediammonium, and the like).

K_(ATP) channel openers defined by Formula III are as follows:

wherein:

-   -   R¹ is selected from the group consisting of hydrogen, lower        alkyl, substituted lower alkyl, and cycloalkyl provided however        that when R¹ is a substituted lower alkyl, then the substituent        does not include an amino group;    -   R^(2a) is hydrogen;    -   R³ is selected from the group consisting of hydrogen, halogen,        lower alkyl, substituted lower alkyl, cycloalkyl and substituted        cycloalkyl provided however that when R³ is a substituted lower        alkyl, then the substituent does not include an amino group;    -   R⁴ is selected from the group consisting of hydrogen, halogen,        lower alkyl, substituted lower alkyl, cycloalkyl and substituted        cycloalkyl provided however that when R⁴ is a substituted lower        alkyl, then the substituent does not include an amino group;    -   and all bioequivalents including salts, prodrugs and isomers        thereof.

In particular embodiments of Formula III, R¹ is a lower alkyl,(preferably ethyl or methyl); R^(2a) is hydrogen; and R³ and R⁴ are eachindependently halogen.

In another embodiment of Formula III, R¹ is methyl; R^(2a) is hydrogen;R³ is selected from the group consisting of hydrogen, halogen, loweralkyl, substituted lower alkyl, cycloalkyl, and substituted cycloalkyl;and R⁴ is chlorine.

Salts of embodiments of the channel openers defined by Formula III maybe prepared from the following: (a) metal hydroxides, preferably alkalimetal hydroxides (e.g., NaOH and KOH) and (b) organic hydroxides,preferably organic compounds which include at least one tertiary amineor at least one quaternary ammonium ion (e.g., diethylaminoethanol,triethylamine, hydroxyethylpyrrolidine, choline andhexamethylhexamethylenediammonium, and the like).

K_(ATP) channel openers defined by Formula IV are as follows:

wherein:

-   -   R¹ is selected from the group consisting of hydrogen, lower        alkyl, substituted lower alkyl, and cycloalkyl provided however        that when R¹ is a substituted lower alkyl, then the substituent        does not include an amino group;    -   R^(2b) is hydrogen;    -   R³ is selected from the group consisting of hydrogen, halogen,        lower alkyl, substituted lower alkyl, cycloalkyl and substituted        cycloalkyl provided however that when R³ is a substituted lower        alkyl, then the substituent does not include an amino group;    -   R⁴ is selected from the group consisting of hydrogen, halogen,        lower alkyl, substituted lower alkyl, cycloalkyl and substituted        cycloalkyl provided however that when R⁴ is a substituted lower        alkyl, then the substituent does not include an amino group;    -   and all bioequivalents including salts, prodrugs and isomers        thereof.

In particular embodiments of Formula IV, R¹ is a lower alkyl,(preferably ethyl or methyl); R^(2b) is hydrogen; and R³ and R⁴ are eachindependently halogen.

In another embodiment of Formula IV, R¹ is methyl; R^(2b) is hydrogen;R³ is selected from the group consisting of hydrogen, halogen, loweralkyl, substituted lower alkyl, cycloalkyl, and substituted cycloalkyl;and R⁴ is chlorine.

Salts of embodiments of the channel openers defined by Formula IV may beprepared from the following: (a) metal hydroxides, preferably alkalimetal hydroxides (e.g., NaOH and KOH) and (b) organic hydroxides,preferably organic compounds which include at least one tertiary amineor at least one quaternary ammonium ion (e.g., diethylaminoethanol,triethylamine, hydroxyethylpyrrolidine, choline andhexamethylhexamethylenediammonium, and the like).

K_(ATP) channel openers defined by Formula V are as follows:

wherein:

-   -   R¹ is selected from the group consisting of optionally        substituted amino, optionally substituted alkyl, optionally        substituted cycloalkyl, optionally substituted heterocyclyl,        optionally substituted heterocyclylalkyl, optionally substituted        aryl, optionally substituted heteroaryl, and optionally        substituted heteroarylalkyl;    -   R^(2a) is selected from the group consisting of hydrogen, and        lower alkyl;    -   X is a 1, 2 or 3 atom chain, wherein each atom is independently        selected from carbon, sulfur and nitrogen, and each atom is        optionally substituted with halogen, hydroxyl, optionally        substituted lower alkyl, optionally substituted lower alkoxy,        optionally substituted cycloalkyl, or optionally substituted        amino;    -   wherein ring B is saturated, monounsaturated, polyunsaturated or        aromatic;    -   wherein at least one of R¹ or a substituent of X includes an        amino group;    -   and all bioequivalents including salts, prodrugs and isomers        thereof.

In particular embodiments of Formula V, X is C(R^(a))C(R^(b)), whereinR^(a) and R^(b) are independently selected from the group consisting ofhydrogen, halogen, optionally substituted lower alkyl, optionallysubstituted cycloalkyl, optionally substituted lower alkoxy, amino,sulfonylamino, aminosulfonyl, sulfonyl, and the like. Preferably R¹includes at least one substituent containing an amino group. In furtherembodiments, R^(a) and R^(b) are independently selected from the groupconsisting of hydroxyl, substituted oxy, substituted thiol, alkylthio,substituted alkylthio, sulfinyl, sulfonyl, substituted sulfinyl,substituted sulfonyl, substituted sulfonylamino, substituted amino,substituted amine, alkylsulfinyl, alkylsulfonyl, alkylsulfonylamino, andthe like. In a preferred embodiment, Ring B does not include anyheteroatoms.

K_(ATP) channel openers defined by Formula VI are as follows:

wherein:

-   -   R¹ is selected from the group consisting of optionally        substituted amino, optionally substituted alkyl, optionally        substituted cycloalkyl, optionally substituted heterocyclyl,        optionally substituted heterocyclylalkyl, optionally substituted        aryl, optionally substituted heteroaryl, and optionally        substituted heteroarylalkyl;    -   R^(2b) is selected from the group consisting of hydrogen and        lower alkyl;    -   X is a 1, 2 or 3 atom chain, wherein each atom is independently        selected from carbon, sulfur and nitrogen, and each atom is        optionally substituted with halogen, hydroxyl, optionally        substituted lower alkyl, optionally substituted lower alkoxy,        optionally substituted cycloalkyl, or optionally substituted        amino;    -   wherein ring B is saturated, monounsaturated, polyunsaturated or        aromatic;    -   wherein at least one of R¹ or a substituent of X includes an        amino group;    -   and all bioequivalents including salts, prodrugs and isomers        thereof.

In particular embodiments of Formula VI, X is C(R^(a))C(R^(b)), whereinR^(a) and R^(b) are independently selected from the group consisting ofhydrogen, halogen, lower alkyl, substituted lower alkyl, cycloalkyl,substituted cycloalkyl, lower alkoxy, substituted lower alkoxy, amino,sulfonylamino, aminosulfonyl, sulfonyl, and the like. In furtherembodiments, R^(a) and R^(b) are independently selected from the groupconsisting of hydroxyl, substituted oxy, substituted thiol, alkylthio,substituted alkylthio, sulfinyl, sulfonyl, substituted sulfinyl,substituted sulfonyl, substituted sulfonylamino, substituted amino,substituted amine, alkylsulfinyl, alkylsulfonyl, alkylsulfonylamino, andthe like. Preferably R¹ includes at least one substituent containing anamino group. In a preferred embodiment, Ring B does not include anyheteroatoms.

K_(ATP) channel openers defined by Formula VII are as follows:

wherein:

-   -   R¹ is selected from the group consisting of optionally        substituted amino, optionally substituted alkyl, optionally        substituted cycloalkyl, optionally substituted heterocyclyl,        optionally substituted heterocyclylalkyl, optionally substituted        aryl, optionally substituted heteroaryl, and optionally        substituted heteroarylalkyl;    -   R^(2a) is selected from the group consisting of hydrogen, lower        alkyl, and substituted lower alkyl;    -   R³ is selected from the group consisting of hydrogen, halogen,        optionally substituted lower alkyl, optionally substituted        amino, optionally substituted cycloalkyl and optionally        substituted aryl;    -   R⁴ is selected from the group consisting of hydrogen, halogen,        optionally substituted lower alkyl, optionally substituted        amino, optionally substituted cycloalkyl and optionally        substituted aryl;    -   wherein at least one of R¹, R³ and R⁴ includes a substituent        containing an amino group;    -   and all bioequivalents including salts, prodrugs and isomers        thereof.

Preferably, R¹ includes a substituent containing an amino group. Inparticular embodiments of Formula VII; R¹ includes an amino substituent,R^(2a) is hydrogen; and R³ and R⁴ are each independently halogen.

In another embodiment of Formula VII, R^(2a) is hydrogen; R³ is selectedfrom the group consisting of hydrogen, halogen, lower alkyl, substitutedlower alkyl, amino, substituted amino, cycloalkyl, and substitutedcycloalkyl; and R⁴ is chlorine.

K_(ATP) channel openers defined by Formula VIII are as follows:

wherein:

-   -   R¹ is selected from the group consisting of optionally        substituted amino, optionally substituted alkyl, optionally        substituted cycloalkyl, optionally substituted heterocyclyl,        optionally substituted heterocyclylalkyl, optionally substituted        aryl, optionally substituted heteroaryl, and optionally        substituted heteroarylalkyl;    -   R^(2b) is selected from the group consisting of hydrogen, lower        alkyl, and substituted lower alkyl;    -   R³ is selected from the group consisting of hydrogen, halogen,        optionally substituted lower alkyl, optionally substituted        amino, optionally substituted cycloalkyl and optionally        substituted aryl;    -   R⁴ is selected from the group consisting of hydrogen, halogen,        optionally substituted lower alkyl, optionally substituted        amino, optionally substituted cycloalkyl and optionally        substituted aryl;    -   wherein at least one of R¹, R³ and R⁴ includes a substituent        containing an amino group;    -   and all bioequivalents including salts, prodrugs and isomers        thereof.

Preferably R¹ includes a substituent containing an amino group. Inparticular embodiments of Formula VIII, R^(2b) is hydrogen; and R³ andR⁴ are each independently halogen.

In another embodiment of Formula VIII, R^(2b) is hydrogen; R³ isselected from the group consisting of hydrogen, halogen, lower alkyl,optionally substituted lower alkyl, optionally substituted amino, andoptionally substituted cycloalkyl; and R⁴ is chlorine.

Unless otherwise indicated, reference in this application to K_(ATP)channel openers should be understood to refer to K_(ATP) channel openersbased upon a salt of one of the compounds described by Formulae I-VIIIand having one or more, and preferably all three, of the followingproperties: (1) opening SURx/Kir6.y potassium channels, wherein x=1, 2Aor 2B and y=1 or 2; (2) binding to the SURx subunit of K_(ATP) channels;and (3) inhibiting glucose induced release of insulin followingadministration of the compound in vivo. Such K_(ATP) channel openerspreferably have the structure of any of the compounds of Formula I-VIII,or more preferably Formula III-IV where ring B or its equivalent doesnot include any heteroatoms. More preferably, the structure isdiazoxide. Structural variants or bioequivalents of any of the compoundsdefined by Formulae I-VIII, such as derivatives, salts, prodrugs orisomers, are also contemplated herein. Specifically, salts of compoundsof Formula I-IV wherein the cation is selected from a cation of analkali metal or an organic compound which includes a tertiary amine or aquaternary ammonium ion. Preferably, when the salt includes an anion ofdiazoxide and a sodium cation, the salt is not in a form suitable forintravenous use. In other embodiments, when the anion is diazoxide in asolution suitable for intravenous use, the cation is not sodium. Inalternate embodiments, in solutions suitable for intravenous use, whenthe cation is sodium, the anion is not an anion of diazoxide. In certainembodiments, when the salt includes an anion of diazoxide and a sodiumcation, the salt is not in liquid form. More preferably, K_(ATP) channelopeners contemplated herein are salts of compounds of Formulae III andIV wherein the cation is selected from sodium, potassium, choline orhexamethyl hexamethylene diammonium. Other K_(ATP) channel openers thatare contemplated for use herein include BPDZ 62, BPDZ 73, NN414, BPDZ154.

Also provided herein are salts of compounds of Formula V-VIII, whereinat least one substituent of the compound of Formulae V-VIII includes anamino group. In another embodiment, the compound of Formula V-VIII formsthe anion of the salt and a monovalent or divalent metal forms thecation. In other embodiments, the cation includes a tertiary amino orquaternary ammonium group.

In vitro analysis of glucose induced release of insulin via K_(ATP)channel openers can be determined using rat islets as provided by DeTullio et al., J. Med. Chem., 46:3342-3353 (2003), or by using humanislets as provided by Björklund et al., Diabetes, 49:1840-1848 (2000).

Provided herein are formulations, such as controlled releasepharmaceutical formulations, of K_(ATP) channel openers andbioequivalents thereof, which include salts of the compounds of FormulaeI-VIII. In one embodiment, the salt can be formulated for controlledrelease following oral administration. Such formulations contain in asingle administration dosage between 10 and 100 mg, between 25 and 100mg, between 100 and 200 mg, between 200 and 300 mg, between 300 and 500mg or between 500 and 2000 mg of the salt of the K_(ATP) channel openersprovided in Formulae I-VIII. In certain embodiments, the dosage of theK_(ATP) channel openers contained in a formulation may be determinedbased on the weight of the subject for which it is to be administered,i.e., the formulation may contain in a single administration dosagebetween 0.1-20 mg of the K_(ATP) channel opener per kg of the subject'sbody weight, or between 0.1-0.5 mg of the K_(ATP) channel opener per kgof the subject's body weight; or between 0.5-1 mg of the K_(ATP) channelopener per kg of the subject's body weight; or between 1-2 mg of theK_(ATP) channel opener per kg of the subject's body weight, or between2-5 mg of the K_(ATP) channel opener per kg of the subject's bodyweight, or between 5-10 mg of the K_(ATP) channel opener per kg of thesubject's body weight, or between 10-15 mg of the K_(ATP) channel openerper kg of the subject's body weight, or between 15-20 mg of the K_(ATP)channel opener per kg of the subject's body weight.

Also provided herein are controlled release pharmaceutical formulationscontaining K_(ATP) channel openers selected from salts of FormulaeI-VIII, which can be obtained by at least one of the following: (a)particle size reduction involving comminution, spray drying, or othermicronising techniques, (b) use of an ion exchange resin, (c) use ofinclusion complexes, for example cyclodextrin, (d) compaction of theK_(ATP) channel opener with a solubilizing agent including a lowviscosity hypromellose, low viscosity methylcellulose or similarlyfunctioning excipient or combinations thereof, (e) associating theK_(ATP) channel opener with a salt prior to formulation, (f) use of asolid dispersion of the K_(ATP) channel opener, (g) use of a selfemulsifying system, (h) addition of one or more surfactants to theformulation, (i) use of nanoparticles, or (j) combinations of theseapproaches.

Further provided herein are controlled release pharmaceuticalformulations containing K_(ATP) channel openers selected from salts ofthe compounds defined by Formulae I-VIII, which include at least onecomponent that substantially inhibits release of the K_(ATP) channelactivator from the formulation until after gastric transit. As usedherein, “substantially inhibits” means less than 15% release, morepreferably at least less than 10% release, or even more preferably atleast less than 5% release of the drug from the formulation duringgastric transport. Release can be measured in a standard USP basedin-vitro gastric dissolution assay in a calibrated dissolutionapparatus. See e.g., U.S. Pharmacopeia, Chapter 711 (2005).

Also provided are oral pharmaceutical formulations of the K_(ATP)channel openers selected from the salts of the compounds of FormulaeI-VIII, which include at least one component that substantially inhibitsrelease of the K_(ATP) channel opener from the formulation until aftergastric transit. Substantial inhibition of drug release during gastrictransit is achieved by inclusion of a component in the formulationselected from the group consisting of: (a) a pH sensitive polymer orco-polymer applied as a compression coating on a tablet, (b) a pHsensitive polymer or co-polymer applied as a thin film on a tablet, (c)a pH sensitive polymer or co-polymer applied as a thin film to anencapsulation system, (d) a pH sensitive polymer or co-polymer appliedto encapsulated microparticles, (e) a non-aqueous-soluble polymer orcopolymer applied as a compression coating on a tablet, (f) anon-aqueous-soluble polymer or co-polymer applied as a thin film on atablet, (g) a non-aqueous soluble polymer applied as a thin film to anencapsulation system, and (h) a non-aqueous soluble polymer applied tomicroparticles, wherein the pH sensitive polymer or co-polymer isresistant to degradation under acid conditions. Alternatively,substantial inhibition of drug release during gastric transport can alsobe achieved by incorporation of the formulation in an osmotic pumpsystem, by use of systems controlled by ion exchange resins, or bycombinations of any of the above approaches.

Also provided herein are controlled release pharmaceutical formulationsof K_(ATP) channel openers selected from salts of the compounds ofFormulae I-VIII, wherein the formulation includes at least one componentthat contributes to sustained release of a K_(ATP) channel opener overan extended period, e.g., over a period of 2-24 hours followingadministration, or over a period of 2-4 hours following administration,or over a period of 4-8 hours following administration, or over a periodof more than 8-24 hours following administration. These formulations arecharacterized in having one of the following components: (a) a pHsensitive polymeric coating, (b) a hydrogel coating, (c) a film coatingthat controls the rate of diffusion of the drug from a coated matrix,(d) an erodable matrix that controls rate of drug release, (e) polymercoated pellets, granules or microparticles of drug which can be furtherencapsulated or compressed into a tablet, (f) an osmotic pump systemcontaining the drug, (g) a compression coated tablet form of the drug,or (h) combinations of any of the approaches of (a)-(f) above.

As used herein, an erodable matrix is the core of a tablet formulationthat, upon exposure to a suitable aqueous environment, begins a processof disintegration which facilitates the release of drug from the matrix.The rate of release of drug from the tablet is controlled both by thesolubility of the drug and the rate of disintegration of the matrix.

The above formulations may further comprise one or more additionalpharmaceutically active agents (other than K_(ATP) channel openersselected from the salts of the compounds of Formulae I-VIII) useful forthe treatment of a condition selected from the group consisting ofobesity, prediabetes, diabetes, hypertension, depression, elevatedcholesterol, fluid retention, other obesity associated co-morbidities,ischemic and reperfusion injury, epilepsy, cognitive impairment,schizophrenia, mania, other psychotic diseases, and the like.

Further provided is a controlled release pharmaceutical formulation of aK_(ATP) channel opener selected from the salts of the compounds ofFormulae I-VIII wherein administration to an obese, overweight orobesity prone subject results in at least one of the following: (a)inhibition of fasting insulin secretion, (b) inhibition of stimulatedinsulin secretion, (c) elevation of energy expenditure, (d) elevation ofbeta oxidation of fat, or (e) inhibition of hyperphagia for about 24hours.

Additionally provided is a controlled release pharmaceutical formulationof a K_(ATP) channel opener selected from the salts of the compounds ofFormulae I-VIII wherein administration to an obese, overweight orobesity prone subject results in at least one of the following: (a)inhibition of fasting insulin secretion, (b) inhibition of glucosestimulated insulin secretion, (c) elevation of energy expenditure, (d)elevation of beta oxidation of fat, or (e) inhibition of hyperphagia forabout 18 hours.

Still further provided is a controlled release pharmaceuticalformulation of a K_(ATP) channel opener selected from the salts of thecompounds of Formulae I-VIII which upon administration to an obese,overweight or obesity prone subject results in at least one of thefollowing: (a) inhibition of fasting insulin secretion, (b) inhibitionof glucose stimulated insulin secretion, (c) elevation of energyexpenditure, (d) elevation of beta oxidation of fat, or (e) inhibitionof hyperphagia for about 24 hours.

Additionally provided is a controlled release pharmaceutical formulationof a K_(ATP) channel opener selected from the salts of the compounds ofFormulae I-VIII that upon administration to an obese, overweight orobesity prone subject results in at least one of the following: (a)inhibition of fasting insulin secretion, (b) inhibition of glucosestimulated insulin secretion, (c) elevation of energy expenditure, (d)elevation of beta oxidation of fat, or (e) inhibition of hyperphagia forabout 18 hours.

Provided herein is a method of treating hypoglycemia, the methodcomprising orally administering to a subject in need thereof, acontrolled release formulation of a K_(ATP) channel opener selected fromthe salts of the compounds of Formulae I-VIII.

Further provided herein is a method of treating obesity associatedco-morbidities in an obese, overweight or obesity prone subject, themethod comprising administering a therapeutically effective amount of asolid oral dosage form of a K_(ATP) channel opener selected from thesalts of the compounds of Formulae I-VIII, or controlled releasepharmaceutical formulation of a K_(ATP) channel opener selected from thesalts of the compounds of Formulae I-VIII. In a preferred embodiment,administration is no more than two times per 24 hours, or once per 24hours.

Yet further provided herein is a method of achieving weight loss in anobese overweight, or obesity prone subject, the method comprisingadministering a therapeutically effective amount of a solid oral dosageform of a K_(ATP) channel opener selected from the salts of thecompounds of Formulae I-VIII or controlled release pharmaceuticalformulation of a K_(ATP) channel opener selected from the salts of thecompounds of Formulae I-VIII. In a preferred embodiment, administrationis no more than two times per 24 hours, or once per 24 hours. The dailydosage administered is preferably between 50 and 180 mg. In certainembodiments, the obese subject has a body mass index greater than 30kg/m², or greater than 35 kg/m², or greater than 40 kg/m², or greaterthan 50 kg/m², or greater than 60 kg/m² at the time the methodcommences.

Also provided is a method of maintaining a weight loss in an obeseoverweight, or obesity prone subject, the method comprisingadministering a therapeutically effective amount of a solid oral dosageform of a K_(ATP) channel opener selected from the salts of thecompounds of Formulae I-VIII or controlled release pharmaceuticalformulation of a K_(ATP) channel opener selected from the salts of thecompounds of Formulae I-VIII. It is preferable to maintain weight in anobese subject once some weight loss has occurred when the alternative isto regain weight. In a preferred embodiment, administration is no morethan two times per 24 hours, or once per 24 hours.

Further provided is a method of elevating energy expenditure in anoverweight, obese or obesity prone subject, the method comprisingadministering an effective amount of a solid oral dosage form of aK_(ATP) channel opener selected from the salts of the compounds ofFormulae I-VIII or controlled release pharmaceutical formulation of aK_(ATP) channel opener selected from the salts of the compounds ofFormulae I-VIII. In a preferred embodiment, administration is no morethan two times per 24 hours, or once per 24 hours. In certainembodiments, the subject has a body mass index greater than 20 kg/m², orgreater than 25 kg/m², or greater than 30 kg/m², or greater than 35kg/m², or greater than 40 kg/m², or greater than 50 kg/m², or greaterthan 60 kg/m² at the time the method commences.

Additionally provided is a method of elevating beta oxidation of fat inan overweight, obese or obesity prone subject, the method comprisingadministering an effective amount of a solid oral dosage form of aK_(ATP) channel opener selected from the salts of the compounds ofFormulae I-VIII or controlled release pharmaceutical formulation of aK_(ATP) channel opener selected from the salts of the compounds ofFormulae I-VIII. In a preferred embodiment, administration is no morethan two times per 24 hours, or once per 24 hours. In certainembodiments, the subject has a body mass index greater than 20 kg/m², orgreater than 25 kg/m², or greater than 30 kg/m², or greater than 35kg/m², or greater than 40 kg/m², or greater than 50 kg/m², or greaterthan 60 kg/m² at the time the method commences.

Yet further provided is a method of reducing visceral fat in anoverweight, obese or obesity prone subject, the method comprisingadministering an effective amount of a solid oral dosage form of aK_(ATP) channel opener selected from the salts of the compounds ofFormulae I-VIII or controlled release pharmaceutical formulation of aK_(ATP) channel opener selected from the salts of the compounds ofFormulae I-VIII. In a preferred embodiment, administration is no morethan two times per 24 hours, or once per 24 hours.

Still further provided is a method of delaying or preventing thetransition to diabetes of a prediabetic subject comprising administeringan effective amount of a K_(ATP) channel opener selected from the saltsof the compounds of Formulae I-VIII or controlled release pharmaceuticalformulation of a K_(ATP) channel opener selected from the salts of thecompounds of Formulae I-VIII. In a preferred embodiment, administrationis no more than two times per 24 hours, or once per 24 hours.

Additionally provided is a method of restoring normal glucose tolerancein a prediabetic subject comprising administering an effective amount ofa K_(ATP) channel opener selected from the salts of the compounds ofFormulae I-VIII or controlled release pharmaceutical formulation of aK_(ATP) channel opener selected from the salts of the compounds ofFormulae I-VIII. In a preferred embodiment, administration is no morethan two times per 24 hours, or once per 24 hours.

Further provided is a method of restoring normal glucose tolerance in adiabetic subject comprising administering an effective amount of aK_(ATP) channel opener selected from the salts of the compounds ofFormulae I-VIII or controlled release pharmaceutical formulation of aK_(ATP) channel opener selected from the salts of the compounds ofFormulae I-VIII. In a preferred embodiment, administration is no morethan two times per 24 hours, or once per 24 hours.

Still further provided is a method of delaying or preventing progressionof diabetes in an subject comprising administering an effective amountof a K_(ATP) channel opener selected from the salts of the compounds ofFormulae I-VIII or controlled release pharmaceutical formulation of aK_(ATP) channel opener selected from the salts of the compounds ofFormulae I-VIII. In a preferred embodiment, administration is no morethan two times per 24 hours, or once per 24 hours.

Also provided is a method to prevent or treat weight gain, impairedglucose tolerance or dyslipidemia associated with the administration ofanti-psychotics to a subject, said method including theco-administration of an effective amount of a K_(ATP) channel openerselected from the salts of the compounds of Formulae I-VIII orcontrolled release pharmaceutical formulation of a K_(ATP) channelopener selected from the salts of the compounds of Formulae I-VIII. In apreferred embodiment, administration is no more than two times per 24hours, or once per 24 hours.

Further provided is a method to treat obesity, or hyperphagia in aPrader-Willi Syndrome patient, a Froelich's Syndrome patient, in a CohenSyndrome patient, in a Summit Syndrome patient, in an Alstrom Syndromepatient, in a Borjeson Syndrome patient or in a Bardet-Biedl Syndromepatient comprising the administration of an effective amount of aK_(ATP) channel opener selected from the salts of the compounds ofFormulae I-VIII or controlled release pharmaceutical formulation of aK_(ATP) channel opener selected from the salts of the compounds ofFormulae I-VIII. In a preferred embodiment, administration is no morethan two times per 24 hours, or once per 24 hours.

Still further provided is a method to treat obesity or elevatedtriglycerides in a patient suffering hyperlipoproteinemia type I, typeII, type III or type IV comprising administering an effective amount ofa K_(ATP) channel opener selected from the salts of the compounds ofFormulae I-VIII or controlled release pharmaceutical formulation of aK_(ATP) channel opener selected from the salts of the compounds ofFormulae I-VIII. In a preferred embodiment, administration is no morethan two times per 24 hours, or once per 24 hours.

Also provided is a method of reducing the incidence of adverse effectsfrom administration of a K_(ATP) channel opener selected from the saltsof the compounds of Formulae I-VIII in the treatment of diseases of asubject achieved by any of the following: (a) use of a dosage form thaton administration reduces C_(max) relative to the current Proglycem®oral suspension or capsule products in order to reduce the incidence ofadverse side effects that are associated with peak drug levels, (b) useof a dosage form that delays release until gastric transit is completein order to reduce the incidence of adverse side effects that areassociated with the release of drug in the stomach, (c) initiatingdosing at subtherapeutic levels and in a stepwise manner increasing dosedaily until the therapeutic dose is achieved wherein the number of stepsis 2 to 10 to reduce the incidence of adverse side effects that occurtransiently at the initiation of treatment, (d) use of the lowesteffective dose to achieve the desired therapeutic effect in order toreduce the incidence of adverse side effects that are dose dependent, or(e) optimizing the timing of administration of dose within the day andrelative to meals.

Further provided is a method of preventing weight gain, dyslipidemia orimpaired glucose tolerance in a subject treated with an anti-psychoticdrug, the method comprising administering a pharmaceutical formulationof a K_(ATP) channel opener selected from the salts of the compounds ofFormulae I-VIII.

Yet further provided is a method of treating weight gain, dyslipidemiaor impaired glucose tolerance in a subject treated with ananti-psychotic drug, the method comprising administering apharmaceutical formulation of a K_(ATP) channel opener selected from thesalts of the compounds of Formulae I-VIII.

Also provided is a method of treating diseases characterized by obesity,hyperphagia, dyslipidemia, or decreased energy expenditure including (a)Prader-Willi Syndrome, (b) Froelich's syndrome, (c) Cohen syndrome, (d)Summit Syndrome, (e) Alstrom Syndrome, (f) Borjesen Syndrome, (g)Bardet-Biedl Syndrome, or (h) hyperlipoproteinemia type I, II, III, andIV comprising administering a pharmaceutical formulation of a K_(ATP)channel opener selected from the salts of the compounds of FormulaeI-VIII.

Further provided is a pharmaceutical formulation of a K_(ATP) channelopener selected from the salts of the compounds of Formulae I-VIIIfurther comprising a pharmaceutically active agent other than theK_(ATP) channel opener. In this formulation, the other pharmaceuticallyactive agent is an agent useful for the treatment of a conditionselected from the group consisting of obesity, prediabetes, diabetes,hypertension, depression, elevated cholesterol, fluid retention, orother obesity associated co-morbidities, ischemic and reperfusioninjury, epilepsy, cognitive impairment, schizophrenia, mania, and otherpsychotic condition.

The formulations containing K_(ATP) channel openers selected from thesalts of the compounds of Formulae I-VIII described herein provide forimproved compliance, efficacy and safety, and for co-formulations withother agents. Included are co-formulations of K_(ATP) channel openersselected from the salts of the compounds of Formulae I-VIII with one ormore additional pharmaceutically active agents that have complementaryor similar activities or targets. Other pharmaceutically active agentsthat can be combined with K_(ATP) channel openers selected from thesalts of the compounds of Formulae I-VIII to treat obesity or tomaintain weight loss in an obesity prone subject include, but are notlimited to: sibutramine, orlistat, phentermine, rimonabant, a diuretic,an antiepileptic, or other pharmaceutical active whose therapeuticutility includes weight loss. It is preferable to maintain weight in anobese subject once some weight loss has occurred when the alternative isto regain weight. Other pharmaceutically active agents that may becombined with K_(ATP) channel openers selected from the salts of thecompounds of Formulae I-VIII to treat type II diabetes, or prediabetesinclude acarbose, miglitol, metformin, repaglinide, nateglinide,rosiglitazone, proglitazone, ramipril, metaglidasen, or any otherpharmaceutical active that improves insulin sensitivity or glucoseutilization or glycemic control where the mode of action is not enhancedinsulin secretion. Other pharmaceutical active agent that can becombined with K_(ATP) channel openers selected from the salts of thecompounds of Formulae I-VIII to treat obesity associated co-morbiditiesinclude a drug active used to lower cholesterol, a drug active used tolower blood pressure, an anti-inflammatory drug that is not a cox-2inhibitor, a drug that is an antidepressant, a drug used to treaturinary incontinence, or other drug routinely used to treat diseaseconditions the incidence of which is elevated in overweight or obesepatients as compared to normal weight subjects including, but notlimited to, drugs to treat atherosclerosis, osteoarthritis, discherniation, degeneration of knees and hips, breast, endometrium,cervical, colon, leukemia and prostate cancers, hyperlipidemia,asthma/reactive airway disease, gallstones, GERD, obstructive sleepapnea, obesity hypoventilation syndrome, recurrent ventral hernias,menstrual irregularity and infertility.

Also provided herein are methods for treating obesity or obesityassociated co-morbidities or other diseases or conditions involvingK_(ATP) channels by co-administration to a subject in need thereof of aneffective amount of any of the compounds according to Formulae I-VIII,or a salt of any of the compounds according to Formulae I-VIII orpharmaceutical formulation thereof, and a drug selected from the groupconsisting of an amphetamine or amphetamine mixture, Sibutramine,Orlistat, Rimonabant, a CB-1 agonist, a 5HT_(2c) receptor agonist, adrug used to treat addiction, a beta adrenergic receptor agonist, an ACCinhibitor, leptin, a leptin analogue, a leptin agonist, a somatostatinagonist, an adiponectin agonist or secretagogue, Amylin, PYY or a PYYanalogue, a ghrelin antagonist, a drug that inhibits gastrointestinallipases or other digestive enzymes, a de-novo lipogenesis inhibitor, adrug that blocks absorption of dietary fat, growth hormone or a growthhormone analogue, a growth hormone secretagogue, a CCK agonist, anoleoylethanolamine receptor agonist, a fatty acid synthase inhibitor, athyroid receptor agonist, a selective androgen receptor modulator, aPPAR agonist, oxyntomodulin, oleoylestrone, a NPY2 receptor antagonist,a NPY5 receptor antagonist, a NPY agonist, a monoamine uptake inhibitor,a MTP inhibitor, a MC4 receptor agonist, a MCH1 receptor antagonist, a5HT-6 antagonist, a histamine-3 antagonist, a glycine analog, a fgf1inhibitor, a DGAT-1 inhibitor, a carboxypeptidase inhibitor, an appetitesuppressant, a non-thiazide diuretic, a drug that lowers cholesterol, adrug that raises HDL cholesterol, a drug that lowers LDL cholesterol, adrug that lowers blood pressure, a drug that is an anti-depressant, adrug that improves insulin sensitivity, a drug that improves glucoseutilization or uptake, a drug that is an anti-epileptic, a drug that isan anti-inflammatory, a drug that is an appetite suppressant, a drugthat lowers circulating triglycerides, a drug that is used to induceweight loss in an overweight or obese individual, and pharmaceuticallyacceptable salts thereof.

Also provided herein are polymorphic forms (i.e., “polymorphs”) of thecompounds of Formulae I-VIII, as exemplified by the X-ray PowerDiffraction (XRPD) patterns shown in any of the figures.

Also provided are polymorphs of salts of diazoxide which includediazoxide and a cation selected from the group consisting of an alkalimetal and a compound comprising a tertiary amine or quaternary ammoniumgroup.

Also provided herein are methods for producing a diazoxide choline salt,which includes suspending diazoxide in a solvent and mixing with acholine salt, adding a co-solvent to the suspension under conditionssufficient to cause formation and precipitation of the diazoxide cholinesalt, and harvesting the precipitate to provide the diazoxide cholinesalt.

Also provided herein are methods of treating obesity or obesity-relatedco-morbidity in an obese subject, wherein the method comprisingadministering to a subject in need thereof an effective amount of acompound of Formula I-VIII.

Also provided herein are methods for treatment of a subject sufferingfrom or at risk for Alzheimer's disease (AD), which methods includeadministration to a subject a therapeutically effective amount of apharmaceutical formulation comprising a compound of any of FormulaeI-VIII as provided herein. In some embodiments, the compound isdiazoxide. Also provided herein are methods for treatment of a subjectsuffering from or at risk for AD, which methods include administrationto a subject a therapeutically effective amount of a salt of a compoundaccording to any of Formulae I-VIII. In some embodiments, the compoundis a salt of diazoxide.

In another embodiment, the invention provides a method for treatinghypoglycemia by administration of an effective amount of apharmaceutical formulation comprising a salt selected from the groupconsisting of a) a salt comprising an anion of a K_(ATP) channel openerselected from the group consisting of Formula I, Formula II, Formula IIIand Formula IV, and a cation selected from the group consisting of analkali metal and a compound comprising a tertiary amine or ammoniumgroup; b) a salt comprising an anion of a K_(ATP) channel openerselected from the group consisting of Formula V, Formula VI, Formula VIIand Formula VIII; and c) a salt comprising a cation of a K_(ATP) channelopener selected from the group consisting of Formula V, Formula VI,Formula VII and Formula VIII, wherein at least one substituent comprisesan amino group.

In further embodiments, the hypoglycemia is selected from the groupconsisting of a) nighttime hypoglycemia, b) hypoglycemia attributable toa defect in insulin secretion, c) attributable to an insulin secretingtumor, and d) drug-induced hypoglycemia.

In the present context, the term “therapeutically effective” or“effective amount” indicates that the materials or amount of material iseffective to prevent, alleviate, or ameliorate one or more symptoms of adisease or medical condition, and/or to prolong the survival of thesubject being treated.

The term “pharmaceutically acceptable” indicates that the identifiedmaterial does not have properties that would cause a reasonably prudentmedical practitioner to avoid administration of the material to apatient, taking into consideration the disease or conditions to betreated and the respective route of administration. For example, it iscommonly required that such a material be essentially sterile, e.g., forinjectibles.

As used herein, the term “composition” refers to a formulation suitablefor administration to an intended animal subject for therapeuticpurposes that contains at least one pharmaceutically active compound andat least one pharmaceutically acceptable carrier or excipient. Otherterms as used herein are defined below.

Adipocyte: An animal connective tissue cell specialized for thesynthesis and storage of fat.

Agonist: A chemical compound that has affinity for and stimulatesphysiological activity at cell receptors normally stimulated bynaturally occurring substances, triggering a biochemical response. Anagonist of a receptor can also be considered an activator of thereceptor.

About: is used herein to mean in quantitative terms plus or minus 10%.

Adipose tissue: Tissue comprised principally of adipocytes.

Adolescent: A person between 10 and 19 years of age.

Adiponectin: A protein hormone produced and secreted exclusively byadipocytes that regulates the metabolism of lipids and glucose.Adiponectin influences the body's response to insulin. Adiponectin alsohas anti-inflammatory effects on the cells lining the walls of bloodvessels.

Alkali metal: refers to elements included in Group I of the periodictable, such as, lithium, sodium, potassium, rubidium, cesium andfrancium.

Amelioration of the symptoms of a particular disorder by administrationof a particular pharmaceutical composition: refers to any lessening,whether permanent or temporary, lasting or transient that can beattributed to or associated with administration of the composition.

Analog: a compound that resembles another in structure but differs by atleast one atom.

Antagonist: A substance that tends to nullify the action of another, asa drug that binds to a cell receptor without eliciting a biologicalresponse when confronted with an agonist for the receptor.

Atherosclerotic Plaque: A buildup of cholesterol and fatty materialwithin a blood vessel due to the effects of atherosclerosis

Bariatric Surgery: A range of surgical procedures which are designed toaid in the management or treatment of obesity and allied diseases.

Beta cell rest: Temporarily placing beta cells in a condition in whichthere is reduced metabolic stress due to suppressed secretion ofinsulin.

Bilaminate: A component of a pharmaceutical dosage form that consists ofthe lamination of two distinct materials.

Bioavailability: Refers to the amount or extent of therapeuticallyactive substance that is released from the drug product and becomesavailable in the body at the intended site of drug action. The amount orextent of drug released can be established by thepharmacokinetic-parameters, such as the area under the blood or plasmadrug concentration-time curve (AUC) and the peak blood or plasmaconcentration (C_(max)) of the drug.

Bioequivalent: Two formulations of the same active substance arebioequivalent when there is no significant difference in the rate andextent to which the active substance becomes available at the site ofdrug action when administered at the same molar dose under similarconditions. “Formulation” in this definition may include the free baseof the active substance or different salts of the active substance.Bioequivalence may be demonstrated through several in vivo and in vitromethods. These methods, in descending order of preference, includepharmacokinetic, pharmacodynamic, clinical and in vitro studies. Inparticular, bioequivalence is demonstrated using pharmacokineticmeasures such as the area under the blood or plasma drugconcentration-time curve (AUC) and the peak blood or plasmaconcentration (C_(max)) of the drug, using statistical criteria.

Cannabinoid Receptor: Receptors in the endocannabinoid (EC) systemassociated with the intake of food and tobacco dependency. Blocking thecannabinoid receptor may reduce dependence on tobacco and the cravingfor food.

Capsule: refers to a softgel, caplet, or any other encapsulated dosageform known to practitioners in the art, or a portion thereof. Softgelrefers a soft gelatin capsule, in agreement with the acceptednomenclature adopted by the SoftGel Association. A softgel is aone-piece, sealed, soft gelatin (or other film-forming material) shellthat contains a solution, a suspension, or a semi-solid paste.

Combination: Refers to any association between or among two or moreitems. The combination can be two or more separate items, such as twocompositions or two collections. It can be a mixture thereof, such as asingle mixture of the two or more items, or any variation thereof.

Composition: Refers to any mixture. It can be a solution, a suspension,liquid, powder, a paste, aqueous, non-aqueous or any combinationthereof.

Compression tablet: Tablet formed by the exertion of pressure to avolume of tablet matrix in a die.

Compression coated tablet: A tablet formed by the addition of a coatingby compression to a compressed core containing the pharmaceuticalactive. As used herein the term “tablet” is intended to mean the same asa compression tablet unless indicated otherwise.

Derivative: A chemical substance derived from another substance bymodification or substitution.

Daily dosage: The total amount of a drug taken in a 24 hour periodwhether taken as a single dose or taken in multiple doses.

Diazoxide: 7-chloro-3-methyl-2H-1,2,4-benzothiadiazine 1,1 dioxide(shown below with its tautomer) with the empirical formula C₈H₇ClN₂O₂Sand a molecular weight of 230.7.

Encapsulation system: A structural feature that contains drug withinsuch as a pharmaceutical capsule. A gel into which drug is incorporatedalso is considered an encapsulation system.

Equivalent amount: An amount of a derivative of a drug that in assays orupon administration to a subject produces an equal effect to a definedamount of the non-derivatized drug.

Fatty acid synthase: The central enzyme of a multienzyme complex thatcatalyses the formation of palmitate from acetylcoenzyme A,malonylcoenzyme A, and NADPH.

Gastric Lipase: An enzyme secreted into the gastrointestinal tract thatcatalyzes the hydrolysis of dietary triglycerides.

Glidant: An inactive component of a pharmaceutical formulation thatprevents caking of the matrix during processing steps.

Hyperinsulinemia: Excessively high blood insulin levels, which isdifferentiated from hyperinsulinism, excessive secretion of insulin bythe pancreatic islets. Hyperinsulinemia may be the result of a varietyof conditions, such as obesity and pregnancy.

Hyperinsulinism: Excessive secretion of insulin by the pancreaticislets.

Hyperlipidemia: A general term for elevated concentrations of any or allof the lipids in the plasma, such as cholesterol, triglycerides andlipoproteins.

Hyperphagia: Ingestion of a greater than optimal quantity of food.

Ingredient of a pharmaceutical composition: Refers to one or morematerials used in the manufacture of a pharmaceutical composition.Ingredient can refer to an active ingredient (an agent) or to othermaterials in the compositions. Ingredients can include water and othersolvents, salts, buffers, surfactants, non-aqueous solvents, andflavorings.

Insulin resistance: A condition in which the tissues of the body arediminished in their ability to respond to insulin.

Ischemic injury: Injury to tissue that results from a low oxygen stateusually due to obstruction of the arterial blood supply or inadequateblood flow leading to hypoxia in the tissue.

Ketoacidosis: Acidosis accompanied by the accumulation of ketone bodies(ketosis) in the body tissue and fluids, as in diabetic acidosis.

Kit: Refers to a packaged combination. A packaged combination canoptionally include a label or labels, instructions and/or reagents foruse with the combination.

Kir: Pore forming subunit of the K_(ATP) channel. Also known as theinwardly rectifying subunit of the K_(ATP) channel. Typically existingas Kir6.x and infrequently as Kir2.x subspecies.

K_(ATP) channel: An ATP sensitive potassium ion channel across the cellmembrane formed by the association of 4 copies of a sulfonylureareceptor and 4 copies of a pore forming subunit Kir. Agonizing thechannel can lead to membrane hyperpolarization.

K_(ATP) channel opener: As used herein refers to a compound of any ofFormulae I-VIII, or a derivative, salt or prodrug thereof, having one ormore or preferably all of the following three properties: (1) openingSURx/Kir6.y potassium channels, where x=1, 2A or 2B and y=1 or 2; (2)binding to the SURx subunit of K_(ATP) channels; and (3) inhibitingglucose induced release of insulin following administration of thecompound in vivo.

Leptin: Product (16 kD) of the ob (obesity) locus. It is found in plasmaof mammals and exerts a hormonal action, which reduces food uptake andincreases energy expenditure.

Lipogenesis: The generation of new lipids, primarily triacylglycerides.It is dependent on the action of multiple distinct enzymes and transportmolecules.

Lipolysis: The breakdown of fat by the coordinated action of multipleenzymes.

Lipoprotein lipase: An enzyme of the hydrolase class that catalyses thereaction of triacyglycerol and water to yield diacylglycerol and a fattyacid anion. The enzyme hydrolyses triacylglycerols in chylomicrons,very-low-density lipoproteins, low-density lipoproteins, anddiacylglycerols.

Lubricant: An inactive component of a pharmaceutical formulation thatprovides for the flow of materials in various processing steps,particularly tableting.

Microparticle: A small particulate formed in the process of developingpharmaceutical formulations that may be coated prior to producing thefinal dosage from.

Obesity: An increase in body weight beyond the limitation of skeletaland physical requirement, as the result of an excessive accumulation offat in the body. Formally defined as having a body mass index greaterthan 30 kg/m2.

Obesity Prone: Subjects who because of genetic predisposition or priorhistory of obesity are at above average risk of becoming obese.

Obesity related co-morbidities: Any disease or condition of animals orhumans that are increased incidence in obese or overweight subjects.Examples of such conditions include hypertension, prediabetes, type 2diabetes, osteoarthritis and cardiovascular conditions.

Osmotically controlled release: A pharmaceutical dosage form in whichthe release of the active drug is principally achieved by the hydrationof a swellable component of the formulation.

Overweight: An subject whose weight is above that which is ideal fortheir height but who fails to meet the criteria for classification asobese. In humans using Body Mass Index (kg/m2) an overweight subjectshas a BMI between 25 and 30.

Oxidation of Fat: A series of reactions involving acyl-coenzyme Acompounds, whereby these undergo beta oxidation and thioclasticcleavage, with the formation of acetyl-coenzyme A; the major pathway offatty acid catabolism in living tissue.

Pharmaceutical composition: Refers to a composition that contains anagent and one or more other ingredients that is formulated foradministration to a subject. An agent refers to an active ingredient ofa pharmaceutical composition. Typically active ingredients are activefor treatment of a disease or condition. For example, agents that can beincluded in pharmaceutical compositions include agents for treatingobesity or diabetes. The pharmaceutically active agent can be referredto as “a pharmaceutical active.”

Pharmaceutical effect: Refers to an effect observed upon administrationof an agent intended for treatment of a disease or disorder or foramelioration of the symptoms thereof.

Pharmacodynamic: An effect mediated by drug action.

Pharmacokinetic: Relating to the absorption, distribution, metabolismand elimination of the drug in the body.

Polymorph: A crystalline form of a compound that exists in at least twocrystalline forms. Polymorphic forms of any given compound are definedby the same chemical formula and/or composition and are as distinct inchemical structure as crystalline structures of two different chemicalcompounds. Such compounds may differ in packing or geometricalarrangement of respective crystalline lattices. The chemical and/orphysical properties or characteristics of the various polymorphs mayvary with each distinct polymorphic form, and may include, but are notlimited to, variations in solubility, melting point, density, hardness,crystal shape, optical and electrical properties, vapor pressure, andstability.

Preadipocyte: A progenitor cell to adipocytes.

Prediabetic: A condition that precedes diagnosis of type II diabetes.Type II diabetes is a form of diabetes mellitus which is characterizedby insulin insensitivity or resistance.

Prodrug: Refers to a compound which, when metabolized, yields thedesired active compound. Typically, the prodrug is inactive, or lessactive than the active compound, but may provide advantageous handling,administration, or metabolic properties. For example, some prodrugs areesters of the active compound; during metabolysis, the ester group iscleaved to yield the active drug. Also, some prodrugs are activatedenzymatically to yield the active compound, or a compound which, uponfurther chemical reaction, yields the active compound.

Prolonged Administration (prolonged basis): Administration of apharmaceutically acceptable formulation of a drug for 7 or more days.Typically, prolonged administration is for at least two weeks,preferably at least one month, and even more preferably at least twomonths (i.e. at least 8 weeks).

Quick dissolving formulation: A pharmaceutical formulation which uponoral administration may release substantially all of the drug activefrom the formulation within 10 minutes.

Release formulation (sustained), (or “sustained release formulation”): Aformulation of pharmaceutical product that, upon administration toanimals, provides for release of the active pharmaceutical over anextended period of time than provided by formulations of the samepharmaceutical active that result in rapid uptake. Similar terms areextended-release, prolonged-release, and slow-release. In all cases, thepreparation, by definition, has a reduced rate of release of activesubstance.

Release formulation (delayed), (or “delayed release formulation”):Delayed-release products are modified-release, but are notextended-release. They involve the release of discrete amount(s) of drugsome time after drug administration, e.g. enteric-coated products, andexhibit a lag time during which little or no absorption occurs.

Release formulation (controlled), (or “controlled release formulation”):A formulation of pharmaceutical product that may include both delay ofrelease of pharmaceutical active upon administration and control ofrelease in the manner described for sustained release.

Salt: The neutral, basic or acid compound formed by the union of an acidor an acid radical and a base or basic radical. Used generally todescribe any ionic compound not containing an oxide or hydroxide ion.

Solid oral dosage form: Pharmaceutical formulations designed for oraladministration including capsules and tablets.

Subject: Refers to animals, including mammals, such as human beings,domesticated animals, and animals of commercial value.

Sulfonylurea receptor: A component of the K_(ATP) channel responsiblefor interaction with sulfonylurea, other K_(ATP) channel antagonists,diazoxide and other K_(ATP) channel agonists.

Tablet: Pharmaceutical dosage form that is produced by forming a volumeof a matrix containing pharmaceutical active and excipients into a sizeand shape suitable for oral administration.

Thermogenesis: The physiological process of heat production in the body.

Threshold Concentration: The minimum circulating concentration of a drugrequired to exert a specific metabolic, physiological or compositionalchange in the body of a treated human or animal.

Treatment: Any manner in which the symptoms of a condition, disorder ordisease or other indication, are ameliorated or otherwise beneficiallyaltered.

Triglyceride: Storage fats of animal and human adipose tissueprincipally consisting of glycerol esters of saturated fatty acids.

Type I diabetes: A chronic condition in which the pancreas makes littleor no insulin because the beta cells have been destroyed.

Uncoupling protein: A family of proteins that allow oxidation inmitochondria to proceed without the usual concomitant phosphorylation toproduce ATP.

Visceral fat: Human adipose tissues principally found below thesubcutaneous fat and muscle layer in the body.

BRIEF DESCRIPTION OF THE FIGURES

FIG. 1 shows UV spectra of the free form diazoxide and the sodium andpotassium salts of diazoxide in acetonitrile.

FIG. 2 shows UV spectra of the free form diazoxide at varying pH.

FIG. 3 shows UV spectra of the free form diazoxide and sodium andpotassium salts of diazoxide in methanol.

FIG. 4A, FIG. 4B, FIG. 4C, and FIG. 4D show X-Ray Powder Diffractionpatterns for (A) free form diazoxide, (B) potassium salt of diazoxidefrom THF, (C) lysine salt of diazoxide from MEK, and (D) sodium salt ofdiazoxide from acetonitrile, respectively.

FIG. 5A, FIG. 5B and FIG. 5C show NMR spectra (DMSO-d6 solvent) for (A)free form diazoxide, (B) potassium salt, and (C) sodium salt,respectively.

FIG. 6A, FIG. 6B and FIG. 6C show X-Ray Powder Diffraction pattern for(A) sodium salt of diazoxide, (B) sodium salt of diazoxide afterslurrying in water, and (C) free form diazoxide, respectively.

FIG. 7 shows DSC spectra for the free form diazoxide (top) and potassiumsalt of diazoxide (bottom). Description: “a” (Integral=−317.56 mJ;normalized=−84.82 Jg⁻¹; Onset=120.81° C.; Peak=121.29° C.); “b”(Integral=−1170.43 mJ; normalized=−154.64 Jg⁻¹; Onset=329.54° C.;Peak=329.21° C.); “c” (Extrap. Peak=355.01° C.; Peak Value=−4.58 mW;normalized=−1.22 Wg⁻¹; Peak=353.53° C.).

FIG. 8 shows TGA spectra for the free form diazoxide (top) and potassiumsalt of diazoxide (bottom).

FIG. 9A, FIG. 9B and FIG. 9C show X-Ray Powder Diffraction pattern for(A) potassium salt of diazoxide, (B) potassium salt of diazoxide afterslurrying in toluene, and (C) potassium salt of diazoxide afterslurrying in toluene for 14 days, respectively.

FIG. 10A, FIG. 10B and FIG. 10C show X-Ray Powder Diffraction patternfor (A) free form diazoxide, (B) choline salt of diazoxide, and (C)hexamethyl hexamethylene diammonium hydroxide salt of diazoxide,respectively.

FIG. 11 shows DSC spectra for the free form diazoxide (top) and cholinesalt of diazoxide (bottom). Description: “a” (Integral=−41.24 mJ;normalized=−8.05 Jg⁻¹; Onset=101.23° C.; Peak=119.29° C.); “b”(Integral=−497.37 mJ; normalized=−97.10 Jg⁻¹; Onset=166.03° C.;Peak=167.27° C.); “c” (Integral=−1167.83 mJ; normalized=−154.29 Jg⁻¹;Onset=329.54° C.; Peak=329.21° C.).

FIG. 12 shows TGA spectra for the free form diazoxide (top) and cholinesalt of diazoxide (bottom).

FIG. 13A, FIG. 13B and FIG. 13C show X-Ray Powder Diffraction patternfor (A) choline salt of diazoxide, (B) choline salt of diazoxide afterslurrying in dichloromethane for 7 days, and (C) choline salt ofdiazoxide after moisture sorption analysis, respectively.

FIG. 14A, FIG. 14B and FIG. 14C show NMR spectra (DMSO-d6 solvent) for(A) free form diazoxide, (B) choline salt, and (C) hexamethylhexamethylene diammonium hydroxide salt of diazoxide, respectively.

FIG. 15A shows overlay XRPD patterns of free form diazoxide, the productof potassium methoxide in methanol, and the product of sodium methoxidein methanol.

FIG. 15B, FIG. 15C and FIG. 15D show the XRPD patterns for product ofpotassium methoxide reaction with diazoxide in methanol, product ofsodium methoxide reaction with diazoxide in methanol, and freeformdiazoxide, respectively.

FIG. 16A and FIG. 16B show XRPD patterns of (A) polymorphic Form A ofthe choline salt of diazoxide, and (B) a mixture of polymorphic forms Aand B of the choline salt of diazoxide, respectively.

FIG. 17A and FIG. 17B show the NMR spectra (DMSO-d6 solvent) for (A)polymorphic Form A of the choline salt of diazoxide, and (B) polymorphicForm B of the choline salt of diazoxide, respectively.

FIG. 18A, FIG. 18B and FIG. 18C show XRPD patterns of (A) polymorphicForm A of the potassium salt of diazoxide, (B) polymorphic Form B of thepotassium salt of diazoxide, and (C) polymorphic Form C of the potassiumsalt of diazoxide, respectively.

FIG. 19A, FIG. 19B, FIG. 19C and FIG. 19D show XRPD patterns of (A)polymorphic Form D of the potassium salt of diazoxide, (B) polymorphicForm E of the potassium salt of diazoxide, (C) polymorphic Form F of thepotassium salt of diazoxide, and (D) polymorphic Form G of the potassiumsalt of diazoxide, respectively.

FIG. 20 shows the DSC spectra of diazoxide choline salt Form A.Description: “a” (Extrap. Peak=120.44° C.; Peak Value=−1.02 mW;normalized=−0.20 Wg⁻¹; Peak=118.63° C.); “b” (Extrap. Peak=167.94° C.;Peak Value=−19.39 mW; normalized=−3.79 Wg⁻¹; Peak=167.27° C.).

FIG. 21 shows the DSC spectra of diazoxide choline salt Form B.Description: “a” (Extrap. Peak=165.05° C.; Peak Value=−3.85 mW;normalized=−0.86 Wg⁻¹; Peak=162.66° C.).

FIG. 22 provides systolic blood pressure (SBP) and diastolic bloodpressure (DBP) for Proglycem Oral Suspension (Proglycem) and DiazoxideCholine Controlled-Release Tablets (DCCRT) at various times followingdose administration (mean±SEM).

FIG. 23 provides pulse rate for Proglycem Oral Suspension (Proglycem)and Diazoxide Choline Controlled-Release Tablets (DCCRT) at varioustimes following dose administration (mean±SEM).

FIG. 24 provides mean plasma diazoxide (±SD) concentrations after a 200mg dose of diazoxide (linear coordinates).

FIG. 25 provides mean plasma diazoxide (±SD) concentrations after a 200mg dose of diazoxide (semilog coordinates).

FIG. 26 provides simulations to steady-state of once daily dosing with200 mg diazoxide.

DETAILED DESCRIPTION OF THE INVENTION

The present invention provides salts of compounds of Formulae I-VIII andmethods for their preparation. Salts of compounds of Formulae I-IV maybe prepared using monovalent alkali metal cations and compounds whichinclude one or more of a tertiary amine or quaternary ammonium moiety.In such salts, the compounds of Formulae I-IV exist in their anionicform. Furthermore, it has been discovered that the selection of asolvent for the preparation of these salts plays an important role insalt formation. Also described herein is the failure to obtain a salt ofdiazoxide from an alkali metal alkoxide using the method described inU.S. Pat. No. 2,986,573.

Compounds of Formulae V-VIII can form both anions and cations, and thussalts can be prepared using a variety of counter ions, including bothanions and cations. Cations of the compounds of Formulae V-VIII can beformed at an amino group, and anions of the compounds of Formulae V-VIIIcan be formed at either an amino group or at the sulfonyl group. Theformation of salts based on compounds of Formulae V-VIII can be done ina variety of solvents, preferably organic solvents.

As discussed herein, two polymorphic forms (i.e., Forms A and B) of thecholine salt of diazoxide have been identified. In summary, both Forms Aand B are anhydrous crystals of diazoxide choline salt. Diazoxidecholine salt Form A can be formed using fast cooling procedures asprovided herein, whereas slow cooling procedures generally favorformation of Form B. Slurry studies shows that Form A readily convertsto Form B. Without wishing to be bound by theory, the slurry studiesindicate that Form B of diazoxide choline salt is the thermodynamicallymore stable form.

Regarding the potassium salt of diazoxide, seven polymorphic forms havebeen identified (i.e., Forms A-G). Diazoxide potassium salt Forms C, D,and F were observed be an acetone solvent, a hemihydrate, and a dioxanesolvent, respectively. Forms A, B, E, and G were not commonly observedduring screening, and elemental analysis suggests that Forms A, B, E andG may be mixtures, have residual solvent present, and/or not be apotassium salt, at least in part. Without wishing to be bound by theory,slurry studies suggest that Form D is the thermodynamically most stablepolymorph of the diazoxide potassium salt polymorphs.

Further provided are pharmaceutical formulations of particular K_(ATP)channel openers of salts of compounds of Formulae I-VIII that whenadministered to subjects achieve novel pharmacodynamic, pharmacokinetic,therapeutic, physiological, and metabolic outcomes. Yet further providedare pharmaceutical formulations, methods of administration and dosing ofparticular K_(ATP) channel openers selected from salts of the compoundsdefined by Formulae I-VIII that achieve therapeutic outcomes whilereducing the incidence of adverse effects.

In particular, pharmaceutical formulations selected from salts ofcompounds defined by Formulae I-VIII and formulated for oraladministration exhibit advantageous properties including: facilitatingconsistency of absorption, pharmacokinetic and pharmacodynamic responsesacross treated patients, contributing to patient compliance andimproving the safety profile of the product, such as by reducing thefrequency of serious adverse effects. Method of treatment of metabolicand other diseases of humans and animals by administering theformulations are also provided.

As shown below, diazoxide and derivatives thereof can exist as protontautomers. Proton tautomers are isomers that differ from each other onlyin the location of a hydrogen atom and a double bond. The hydrogen atomand double bond switch locations between a carbon atom and a heteroatom,such as for example N. Thus, when the substituent on the nitrogen ishydrogen, the two isomeric chemical structures may be usedinterchangeably.

The particular K_(ATP) channel openers that can be used in the inventionformulations include salts of any of the compounds within Formulae I toVIII. Exemplary compounds which have been previously reported includediazoxide, BPDZ 62, BPDZ 73, NN414 and BPDZ 154 (see, for example, Schouet al., Bioorg. Med. Chem., 13, 141-155 (2005)). Compound BPDZ 154 alsois an effective K_(ATP) channel activator in patients withhyperinsulinism and in patients with pancreatic insulinoma. Thesynthesis of BPDZ compound is provided in Cosgrove et al., J. Clin.Endocrinol. Metab., 87, 4860-4868 (2002).

Channel openers demonstrating decreased activity in the inhibition ofinsulin release and increased activity in vascular smooth muscle tissuehave been previously reported and include analogs of diazoxide such as,for example, 3-isopropylamino-7-methoxy-4H-1,2,4,-benzothiadiazine1,1-dioxide, (a selective Kir6.2/SUR1 channel opener; see Dabrowski etal., Diabetes, 51, 1896-1906 (2002), and 2-alkyl substituted diazoxides(see, for example, Ouedraogo et al., Biol. Chem., 383, 1759-1768(2002)). The 2-alkyl substituted diazoxides generally do not function astraditional potassium channel activators, but instead show potential asCa²⁺ blockers.

Other diazoxide analogs which have been previously reported includedescribed in Schou et al., Bioorg. Med. Chem., 13, 141-155 (2005), areshown below.

Diazoxide analogs having different alkyl substituents at the 3 positionof the molecule (identified as R³ shown below) are described inBertolino et al., Receptors and Channels, 1, 267-278 (1993).

K_(ATP) channel activity of salts of the compounds of Formulae I-VIIIand related compounds can be measured by membrane potential studies asdescribed in Schou et al., Bioorg. Med. Chem., 13, 141-155 (2005) andDabrowski, et al., Diabetes, 51, 1896-1906 (2002).

Measurement of the inhibition of glucose-stimulated insulin release fromβTC6 cells is described in Schou et al., Bioorg. Med. Chem., 13, 141-155(2005). The ability of particular K_(ATP) channel openers to inhibitrelease of insulin from incubated rat pancreatic islets can be performedas described by Ouedraogo et al., Biol. Chem., 383, 1759-1768 (2002).

Activation of recombinant K_(ATP) channels by K_(ATP) channel openerscan be examined by monitoring macroscopic currents of inside-outmembrane patches from Xenopus oocytes co-expressing Kir6.2 and eitherSUR1, SUR2A or SUR2B. SUR expressing membranes can be prepared by knownmethods. See, for example, Dabrowski et al., Diabetes, 51, 1896-1906(2002).

Binding experiments can be used to determine the ability of K_(ATP)channel openers to bind SUR1, SUR2A and SUR2B. See, for example,Schwanstecher et al., EMBO J., 17, 5529-5535 (1998).

Preparation of SUR1 and SUR2A chimeras, as described by Babenko et al.,allows for comparison of pharmacologic profiles (i.e. sulfonylsensitivity and responsiveness to diazoxide or other potassium channelopeners) of the SUR1/Kir6.2 and SUR2A/Kir6.2 potassium channels. SeeBabenko et al., J. Biol. Chem., 275(2), 717-720 (2000). The cloning of asulfonylurea receptor and an inwardly rectifying K⁺ channel is describedby Isomoto et al., J. Biol. Chem., 271 (40), 24321-24324 (1996); D'hahanet al., PNAS, 96(21), 12162-12167 (1999).

Differences between the human SUR1 and human SUR2 genes are describedand shown in Aguilar-Bryan et al., Physiological Review, 78(1):227-245(1998).

“Halo” and “halogen” refer to all halogens, that is, chloro (Cl), fluoro(F), bromo (Br), or iodo (I).

“Hydroxyl” and “hydroxy” refer to the group —OH.

“Substituted oxy” refers to the group —OR^(aa), where R^(aa) can bealkyl, substituted alkyl, acyl, substituted acyl, aryl, substitutedaryl, heteroaryl, substituted heteroaryl, aralkyl, substituted aralkyl,cycloalkyl, substituted cycloalkyl, heterocyclyl, or substitutedheterocyclyl.

“Substituted thiol” refers to the group —SR^(bb), where R^(bb) can bealkyl, substituted alkyl, acyl, substituted acyl, aryl, substitutedaryl, heteroaryl, substituted heteroaryl, aralkyl, substituted aralkyl,cycloalkyl, substituted cycloalkyl, heterocyclyl, or substitutedheterocyclyl.

“Alkyl” refers to an alkane-derived radical containing from 1 to 10,preferably 1 to 6, more preferably 1-4, yet more preferably 1-2, carbonatoms. Alkyl includes straight chain alkyl, branched alkyl andcycloalkyl, such as methyl, ethyl, propyl, isopropyl, butyl, t-butyl,and the like. The alkyl group can be attached at any available point toproduce a stable compound. An “alkylene” is a divalent alkyl.

A “substituted alkyl” is an alkyl group independently substituted with 1or more, e.g., 1, 2, or 3, groups or substituents such as halo, hydroxy,optionally substituted alkoxy, optionally substituted alkylthio,alkylsulfinyl, alkylsulfonyl, optionally substituted amino, optionallysubstituted amido, amidino, urea optionally substituted with alkyl,aminosulfonyl optionally N-mono- or N,N-di-substituted with alkyl,alkylsulfonylamino, carboxyl, heterocycle, substituted heterocycle,nitro, cyano, thiol, sulfonylamino or the like attached at any availablepoint to produce a stable compound. In particular, “fluoro substituted”refers to substitution by 1 or more, e.g., 1, 2, or 3 fluorine atoms.“Optionally fluoro substituted” means that substitution, if present, isfluoro. The term “optionally substituted” as used herein means thatsubstitution may, but need not, be present.

“Lower alkyl” refers to an alkyl group having 1-6 carbon atoms.

A “substituted lower alkyl” is a lower alkyl which is substituted with 1or more, e.g., 1, 2, or 3, groups or substituents, as defined above,attached at any available point to produce a stable compound.

“Cycloalkyl” refers to saturated or unsaturated, non-aromaticmonocyclic, bicyclic or tricyclic carbon ring systems of 3-8, morepreferably 3-6, ring members per ring, such as cyclopropyl, cyclopentyl,cyclohexyl, adamantyl, and the like. “Cycloalkylene” is a divalentcycloalkyl.

“Substituted cycloalkyl” refers to saturated or unsaturated,non-aromatic monocyclic, bicyclic or tricyclic carbon ring systems of3-8, more preferably 3-6, ring members per ring, such as cyclopropyl,cyclopentyl, cyclohexyl, adamantyl, and the like independentlysubstituted with 1 or more, e.g., 1, 2, or 3, groups or substituentssuch as halo, hydroxy, optionally substituted alkoxy, optionallysubstituted alkylthio, alkylsulfinyl, alkylsulfonyl, optionallysubstituted amino, optionally substituted amido, amidino, ureaoptionally substituted with alkyl, aminosulfonyl optionally N-mono- orN,N-di-substituted with alkyl, alkylsulfonylamino, carboxyl,heterocycle, substituted heterocycle, nitro, cyano, thiol, sulfonylaminoor the like attached at any available point to produce a stablecompound.

“Aryl” alone or in combination means phenyl or naphthyl optionallycarbocyclic fused with a cycloalkyl of preferably 5-7, more preferably5-6, ring members.

“Substituted aryl” refers to an aryl group as defined aboveindependently substituted with 1 or more, e.g., 1, 2, or 3, groups orsubstituents such as halo, hydroxy, optionally substituted alkoxy,optionally substituted alkylthio, alkylsulfinyl, alkylsulfonyl,optionally substituted amino, optionally substituted amido, amidino,urea optionally substituted with alkyl, aminosulfonyl optionally N-mono-or N,N-di-substituted with alkyl, alkylsulfonylamino, carboxyl,heterocycle, substituted heterocycle, nitro, cyano, thiol, sulfonylaminoor the like attached at any available point to produce a stablecompound.

“Alkoxy” denotes the group —OR^(cc), where R^(cc) is alkyl. “Loweralkoxy” denotes the group —OR^(ccc), where R^(ccc) is lower alkyl

“Substituted alkoxy” denotes the group —OR^(dd), where R^(dd) issubstituted alkyl. “Substituted lower alkoxy” denotes the group—OR^(ddd), where R^(ddd) is substituted lower alkyl.

“Alkylthio” or “thioalkoxy” refers to the group —S-R^(ee), where R^(ee)is alkyl.

“Substituted alkylthio” or “substituted thioalkoxy” refers to the group—S—R, where R is substituted alkyl.

“Sulfinyl” denotes the group —S(O)—.

“Sulfonyl” denotes the group —S(O)₂—.

“Substituted sulfinyl” denotes the group —S(O)—R^(ff), where R^(ff) isalkyl, substituted alkyl, cycloalkyl, substituted cycloalkyl,cycloalkylalkyl, substituted cycloalkylalkyl, heterocyclyl, substitutedheterocyclyl, heterocyclylalkyl, substituted hetereocyclylalkyl, aryl,substituted aryl, heteroaryl, substituted heteroaryl, heteroaralkyl,substituted heteroaralkyl, aralkyl or substituted aralkyl.

“Substituted sulfonyl” denotes the group —S(O)₂R^(gg), where R^(gg) isalkyl, substituted alkyl, cycloalkyl, substituted cycloalkyl,cycloalkylalkyl, substituted cycloalkylalkyl, heterocyclyl, substitutedheterocyclyl, heterocyclylalkyl, substituted hetereocyclylalkyl, aryl,substituted aryl, heteroaryl, substituted heteroaryl, heteroaralkyl,substituted heteroaralkyl, aralkyl or substituted aralkyl.

“Sulfonylamino” denotes the group —S(O)₂NR^(hh)— where R^(hh) ishydrogen or alkyl.

“Substituted sulfonylamino” denotes the group —S(O)₂NR^(ii)—R^(jj),where R^(ii) is hydrogen or optionally substituted alkyl, and R^(jj) isalkyl, substituted alkyl, cycloalkyl, substituted cycloalkyl,heterocyclyl, substituted heterocyclyl, aryl, substituted aryl,heteroaryl, substituted heteroaryl, heteroaralkyl, substitutedheteroaralkyl, aralkyl or substituted aralkyl.

“Amino” or “amine” denotes the group —NH₂. A “divalent amine” denotesthe group —NH—. A “substituted divalent amine” denotes the group—NR^(kk)— wherein R^(kk) is alkyl, substituted alkyl, aryl, substitutedaryl, heteroaryl, substituted heteroaryl, acyl, substituted acyl,sulfonyl or substituted sulfonyl.

“Substituted amino” or “substituted amine” denotes the group—NR^(mm)R^(nn), wherein R^(mm) and R^(nn) are independently hydrogen,alkyl, substituted alkyl, aryl, substituted aryl, heteroaryl,substituted heteroaryl, acyl, substituted acyl, sulfonyl, substitutedsulfonyl, or cycloalkyl provided, however, that at least one of R^(mm)and R^(nn) is not hydrogen. R^(mm)R^(nn) in combination with thenitrogen may form an optionally substituted heterocyclic or heteroarylring.

“Alkylsulfinyl” denotes the group —S(O)R^(oo), wherein R^(oo) isoptionally substituted alkyl.

“Alkylsulfonyl” denotes the group —S(O)₂R^(pp), wherein R^(pp) isoptionally substituted alkyl.

“Alkylsulfonylamino” denotes the group —NR^(qq)S(O)₂R^(rr), whereinR^(rr) is optionally substituted alkyl, and R^(qq) is hydrogen or alkyl.

A “primary amino substituent” denotes the group —NH₂.

A “secondary amino substituent” denotes the group —NHR^(ss), whereinR^(ss) is alkyl, substituted alkyl, aryl, substituted aryl, heteroaryl,substituted heteroaryl, acyl, substituted acyl, sulfonyl, substitutedsulfonyl, or cycloalkyl.

A “tertiary amino substituent” denotes the group —NR^(ss)R^(tt), whereinR^(as) and R^(tt) are independently alkyl, substituted alkyl, aryl,substituted aryl, heteroaryl, substituted heteroaryl, acyl, substitutedacyl, sulfonyl, substituted sulfonyl, or cycloalkyl.

“Quaternary ammonium substituent” denotes the group—N⁺R^(ss)R^(tt)R^(uu), wherein R^(ss), R^(tt) and R^(uu) areindependently alkyl, substituted alkyl, aryl, substituted aryl,heteroaryl, substituted heteroaryl, acyl, substituted acyl, sulfonyl,substituted sulfonyl, or cycloalkyl.

“Heteroaryl” means a monocyclic aromatic ring structure containing 5 or6 ring atoms, or a bicyclic aromatic group having 8 to 10 atoms,containing one or more, preferably 1-4, more preferably 1-3, even morepreferably 1-2, heteroatoms independently selected from the groupconsisting of O, S, and N. Heteroaryl is also intended to includeoxidized S or N, such as sulfinyl, sulfonyl and N-oxide of a tertiaryring nitrogen. A carbon or nitrogen atom is the point of attachment ofthe heteroaryl ring structure such that a stable aromatic ring isretained. Examples of heteroaryl groups are pyridinyl, pyridazinyl,pyrazinyl, quinaoxalyl, indolizinyl, benzo[b]thienyl, quinazolinyl,purinyl, indolyl, quinolinyl, pyrimidinyl, pyrrolyl, oxazolyl,thiazolyl, thienyl, isoxazolyl, oxathiadiazolyl, isothiazolyl,tetrazolyl, imidazolyl, triazinyl, furanyl, benzofuryl, indolyl, and thelike. “Heteroarylene” means a divalent heteroaryl.

“Heterocycle” or “heterocyclyl” means a saturated or unsaturated,non-aromatic carbocyclic group having a single ring or multiplecondensed rings, e.g. a cycloalkyl group having from 5 to 10 atoms inwhich from 1 to 3 carbon atoms in a ring are replaced by heteroatoms,such as O, S, N, and are optionally fused with benzo or heteroaryl of5-6 ring members and/or are optionally substituted. Heterocyclyl isintended to include oxidized S or N, such as sulfinyl, sulfonyl andN-oxide of a tertiary ring nitrogen. Examples of heterocycle orheterocyclyl groups are morpholino, tetrahydrofuranyl, dihydropyridinyl,piperidinyl, pyrrolidinyl, piperazinyl, dihydrobenzofuryl,dihydroindolyl, and the like.

“Heterocyclylalkyl” refers to the group —R-Het where Het is aheterocycle group and R is an alkylene group.

A “substituted heteroaryl,” “substituted heterocyclyl,” or “substitutedheterocyclylalkyl” is a heteroaryl, heterocyclyl, or heterocyclylalkyl,respectively, independently substituted with 1 or more, e.g., 1, 2, or3, groups or substituents such as halogen, hydroxy, optionallysubstituted alkoxy, optionally substituted alkylthio, alkylsulfinyl,alkylsulfonyl, acyloxy, optionally substituted aryl, optionallysubstituted aryloxy, optionally substituted heteroaryloxy, optionallysubstituted amino, optionally substituted amido, amidino, ureaoptionally substituted with alkyl, aryl, heteroaryl or heterocyclylgroups, aminosulfonyl optionally N-mono- or N,N-di-substituted withalkyl, aryl or heteroaryl groups, alkylsulfonylamino, arylsulfonylamino,heteroarylsulfonylamino, alkylcarbonylamino, arylcarbonylamino,heteroarylcarbonylamino, carboxyl, heterocycle, substituted heterocycle,heteroaryl, substituted heteroaryl, nitro, cyano, thiol, sulfonylamino,optionally substituted alkyl, optionally substituted alkenyl, oroptionally substituted alkynyl, attached at any available point toproduce a stable compound.

“Amido” denotes the group —C(O)NH₂. “Substituted amido” denotes thegroup —C(O)NR^(k)R^(l), wherein R^(k) and R^(l) are independentlyhydrogen, lower alkyl, substituted lower alkyl, aryl, substituted aryl,heteroaryl, or substituted heteroaryl, provided, however, that at leastone of R^(k) and R^(l) is not hydrogen. R^(k)R^(l) in combination withthe nitrogen may form an optionally substituted heterocyclic orheteroaryl ring.

“Amidino” denotes the group —C(═NR^(m))NR^(n)R^(o), wherein R^(m),R^(n), and R^(o) are independently hydrogen or optionally substitutedlower alkyl.

“Acyloxy” denotes the group —OC(O)R^(h), where R^(h) is hydrogen, alkyl,substituted alkyl, cycloalkyl, substituted cycloalkyl, heterocyclyl,substituted heterocyclyl, aryl, substituted aryl, heteroaryl,substituted heteroaryl and the like.

“Aryloxy” denotes the group —OAr, where Ar is an aryl, or substitutedaryl, group. “Heteroaryloxy” denotes groups —OHet, wherein Het is anoptionally substituted heteroaryl group.

“Arylsulfonylamino” denotes the group —NR^(q)S(O)₂R^(s), wherein R^(s)is optionally substituted aryl, and R^(q) is hydrogen or lower alkyl.“Heteroarylsulfonylamino” denotes the group —NR^(q)S(O)₂R^(t), whereinR^(t) is optionally substituted heteroaryl, and R^(q) is hydrogen orlower alkyl.

“Alkylcarbonylamino” denotes the group —NR^(q)C(O)R^(p), wherein R^(P)is optionally substituted alkyl, and R^(q) is hydrogen or lower alkyl.

“Arylcarbonylamino” denotes the group —NR^(q)C(O)R^(s), wherein R^(s) isoptionally substituted aryl, and R^(q) is hydrogen or lower alkyl.

“Heteroarylcarbonylamino” denotes the group —NR^(q)C(O)R^(t), whereinR^(t) is optionally substituted aryl, and R^(q) is hydrogen or loweralkyl.

Pharmaceutical formulations containing K_(ATP) channel openers caninclude the free base of a compound defined by any of Formulae I-VIII,or a salt thereof. Salts of the compounds of Formulae I-VIII as providedherein may have one or more of the following characteristics: (1)stability in solution during synthesis and formulation, (2) stability ina solid state, (3) compatibility with excipients used in the manufactureof tablet formulations, (4) quantitatively yield the K_(ATP) channelopener upon exposure to simulated or actual gastric and duodenalconditions, (5) release K_(ATP) channel opener from sufficiently smallparticles that are readily dissolved and absorbed, (6) provide, whenincorporated into a pharmaceutical formulation, for absorption ofgreater than 80% of the administered dose, (7) present no elevatedtoxicological risk as compared to the free base of the K_(ATP) channelopener, (8) can be formulated into acceptable pharmaceuticalformulations to treat obesity and other diseases of humans, (9) areacceptable to the FDA as the basis of a drug product, (10) can berecrystallized to improve purity, (11) can be used to form co-crystalsof two or more salts of the K_(ATP) channel opener, (12) have limitedhygroscopicity to improve stability, (13) synthetic and crystallizationconditions under which the salt is formed can be varied resulting indifferent crystal structures (polymorphs) can be controlled in thesynthesis of the salt, or (14) have improved solubility as compared tothe free base in aqueous systems at physiological pH values.

The K_(ATP) channel openers provided in Formulae I-VIII are preferablyformulated as pharmaceutically acceptable salts. Pharmaceuticallyacceptable salts are non-toxic salts in the amounts and concentrationsat which they are administered. The preparation of such salts canfacilitate the pharmacological use by altering the physicalcharacteristics of a compound without preventing it from exerting itsphysiological effect. Useful alterations in physical properties includelowering the melting point to facilitate transmucosal administration andincreasing the solubility to facilitate administering lower effectivedoses of the drug.

Salts of the compounds of Formulae I-IV can include metal cations,preferably alkali metal cations, such as for example, sodium orpotassium. Cations can be selected from any group I alkali metal.Divalent metals cations, such as alkaline earth metals (e.g., magnesium,calcium and the like), have not been found to be useful for saltformation with the compounds of Formulae I-IV.

Salts of the compounds of Formulae I-IV which include alkali metalcations can be prepared by reacting the compounds of Formulae I-IV withan alkali metal hydroxide or alkali metal alkoxide, such as for example,NaOH, KOH or NaOCH3, in a variety of solvents which may be selected fromlow molecular weight ketones (e.g., acetone, methyl ethyl ketone),tetrahydrofuran (THF), dimethylformamide (DMF), andn-methylpyrrolidinone, and the like. Surprisingly, salt formation withan alkali metal hydroxide or alkoxide is not observed when an alcohol,particularly a lower alcohol such as for example methanol or ethanol, isused as the solvent. This result was confirmed by both X-Ray PowderDiffraction and NMR, and is contrary to the disclosure of U.S. Pat. No.2,986,573, which purports to describe formation of diazoxide salts inalcohol.

The compounds of Formulae I-IV can also form salts with organic cationsthat include at least one tertiary amine or ammonium cation. Organiccation compounds can be monovalent, divalent, trivalent and tetravalentby inclusion of one, two, three or four tertiary amine or ammonium ionswithin the compound, respectively. When a multivalent compound is used,the tertiary amine or quaternary ammonium moieties are preferablyseparated by a chain of at least 4 atoms, more preferably by a chain ofat least 6 atoms, such as for example, hexamethyl hexamethylenediammonium dihydroxide, wherein the quaternary ammonium moieties areseparated by —(CH₂)₆—. Primary and secondary amines do not toeffectively form salts with the compounds of Formulae I-IV.

Salts of the compounds of Formulae I-IV can be prepared by reacting thecompounds of Formulae I-IV with compounds that include at least onetertiary amine or quaternary ammonium ion (e.g., choline hydroxide,hexamethylhexamethylene diammonium dihydroxide) in a solvent selectedfrom low molecular weight ketones (e.g., acetone, methyl ethyl ketone),tetrahydrofuran, dimethylformamide, and n-methyl pyrrolidinone. As withthe preparation of salts from alkali metal hydroxides, amine andammonium containing compounds do not form salts when the solvent is analcohol.

Pharmaceutically acceptable salts of the compounds of Formulae I-IV canalso include basic addition salts such as those containing benzathine,chloroprocaine, choline, diethylamino-ethanol, hydroxyethyl pyrrolidine,ammonium, tetrapropylammonium, tetrabutylphosphonium, hexamethyldiammonium, methyldiethanamine, triethylamine, meglumine, and procaine,and can be prepared using the appropriate corresponding bases.

Preferred basic addition salts of the compounds of Formulae I-IV caninclude those containing hexamethyl hexamethylene diammonium, choline,sodium, potassium, methyldiethyl amine, triethylamine,diethylamino-ethanol, hydroxyethyl pyrrolidine, tetrapropylammonium andtetrabutylphosphonium ions.

Preferred basic addition salts of the compounds of Formulae I-IV can beprepared using hexamethyl hexamethylene diammonium dihydroxide, cholinehydroxide, sodium hydroxide, sodium methoxide, potassium hydroxide,potassium methoxide, ammonium hydroxide, tetrapropylammonium hydroxide,and tetrabutylphosphonium hydroxide. The basic addition salts can beseparated into inorganic salts (e.g., sodium, potassium and the like)and organic salts (e.g., choline, hexamethyl hexamethylene diammoniumhydroxide, and the like).

The compounds of Formulae V-VIII have the unique property of being ableto form both anions and cations. In basic media, the compounds ofFormulae V-VIII typically form anions. Anions can be formed at either anamino or substituted amino substituent, or at the sulfonyl group. Inacidic media, the compounds of Formulae V-VIII generally form cations byprotonation of an amino group, thereby forming an ammonium moiety.

Salts of the anions of compounds of Formulae V-VIII can include metalcations, including monovalent metal cations of any group I alkali metal(e.g., sodium, potassium, and the like), divalent metal cations of anygroup II alkaline earth metal (e.g., calcium, magnesium, and the like),and aluminum cations.

Salts of the compounds of Formulae V-VIII which include metal cationscan be prepared by reacting the compounds of Formulae V-VIII with aalkali or alkaline earth metal hydroxides or alkoxides, such as forexample, sodium hydroxide or sodium methoxide, in an organic solvent,such as for example lower alcohols, low molecular weight ketones (e.g.,acetone, methyl ethyl ketone, and the like), tetrahydrofuran,dimethylformamide, and n-methylpyrrolidinone, and the like.

Salts of the compounds of Formulae V-VIII, may include organic orinorganic counter ions, including but not limited to, acetate,acetonide, acetyl, adipate, aspartate, besylate, biacetate, bitartrate,bromide, butoxide, butyrate, calcium, camsylate, caproate, carbonate,citrate, cypionate, decanoate, diacetate, dimegulumine, dinitrate,dipotassium, dipropionate, disodium, disulfide, edisylate, enanthate,estolate, etabonate, ethylsuccinate, fumarate, furoate, gluceptate,gluconate, hexacetonide, hippurate, hyclate, hydrobromide,hydrochloride, isethionate, lactobionate, malate, maleate, meglumine,methylbromide, methylsulfate, metrizoate, nafate, napsylate, nitrate,oleate, palmitate, pamoate, phenpropionate, phosphate, pivalate,polistirex, polygalacturonate, probutate, propionate, saccharate, sodiumglycinate, sodium phosphate, sodium succinate, stearate, succinate,sulfate, sulfonate, sulfosalicylate, tartrate, tebutate, terephalate,terephthalate, tosylate, triflutate, trihydrate, trisilicate,tromethamine, valerate, or xinafoate. Preferred organic cations includecompounds having tertiary amines or quaternary ammonium groups.

Other, pharmaceutically acceptable salts of the compounds of FormulaeV-VIII include acid addition salts such as those containing sulfate,chloride, hydrochloride, fumarate, maleate, phosphate, sulfamate,acetate, citrate, lactate, tartrate, methanesulfonate, ethanesulfonate,benzenesulfonate, p-toluene sulfonate, cyclohexylsulfamate and quinate.Pharmaceutically acceptable salts of the compounds of Formulae V-VIIIcan be obtained from acids such as hydrochloric acid, maleic acid,sulfuric acid, phosphoric acid, sulfamic acid, acetic acid, citric acid,lactic acid, tartaric acid, malonic acid, methanesulfonic acid,ethanesulfonic acid, benzenesulfonic acid, p-toluenesulfonic acid,cyclohexylsulfamic acid, fumaric acid, and quinic acid.

Pharmaceutically acceptable salts of the compounds of Formulae V-VIIIalso include basic addition salts such as those containing benzathine,chloroprocaine, choline, diethanolamine, ethylenediamine, meglumine,procaine, aluminum, calcium, lithium, magnesium, potassium, sodium,ammonium, alkylamine, and zinc, when acidic functional groups, such ascarboxylic acid or phenol are present. For example, see Remington'sPharmaceutical Sciences, 19th ed., Mack Publishing Co., Easton, Pa.,Vol. 2, p. 1457, 1995. Such salts of the compounds of Formulae V-VIIIcan be prepared using the appropriate corresponding bases.

Salts of the compounds of Formulae V-VIII can be prepared, for example,by dissolving the free-base form of a compound in a suitable solvent,such as an aqueous or aqueous-alcohol in solution containing theappropriate acid and then isolated by evaporating the solution. Inanother example, a salt is prepared by reacting the free base and acidin an organic solvent.

The salts of the compounds of Formulae V-VIII may be present as acomplex. Examples of complexes include 8-chlorotheophylline complex(analogous to, e.g., dimenhydrinate: diphenhydramine8-chlorotheophylline (1:1) complex; Dramamine) and various cyclodextrininclusion complexes.

Solvents useful in the preparation of pharmaceutically acceptable saltsof the compounds of Formulae V-VIII include organic solvents, such asfor example, acetonitrile, acetone, alcohols (e.g., methanol, ethanoland isopropanol), tetrahydrofuran, methyl ethyl ketone (MEK), ethers(e.g., diethyl ether), benzene, toluene, xylenes, dimethylformamide(DMF), and N-methylpyrrolidinone (NMP), and the like. Preferably, thesolvents are selected from acetonitrile and MEK.

The salts of compounds of Formulae V-VIII may be present as a complex.Examples of complexes include 8-chlorotheophylline complex (analogousto, e.g., dimenhydrinate: diphenhydramine 8-chlorotheophylline (1:1)complex; Dramamine) and various cyclodextrin inclusion complexes.

Formulations of salts of the compounds of Formulae I-VIII providedherein exhibit at least one, or preferably some or even more preferably,all the following characteristics: (1) they are stable at ambienttemperatures for a minimum of one year; (2) they provide for ease oforal administration; (3) they facilitate patient compliance with dosing;(4) upon administration, they consistently facilitate high levels ofabsorption of the pharmaceutical active; (5) upon once or twice dailyoral administration they allow release of the K_(ATP) channel openerover a sustained time frame such that the circulating concentration ofthe K_(ATP) channel opener or its metabolically active metabolites doesnot fall below a therapeutically effective concentration; (6) theyachieve these results independent of the pH of the gastrointestinaltract of treated subjects, and (7) they delay release until gastrictransit is complete or nearly complete.

Formulations designed for oral administration of the salts of thecompounds of Formulae I-VIII can be provided, for example, as capsules,tablets, or as quick dissolve tablets or films. Capsule or tabletformulations include a number of distinguishing components. One is acomponent to improve absorption of the K_(ATP) channel opener. Anothersustains release of the drug over more than 2 hours. A third delayssubstantial release of the drug until gastric transit is completed.

Oral administration formulations of the salts of the compounds ofFormulae I-VIII can also be provided, for example, as oral suspensions,oral solutions, encapsulated oral suspensions, and encapsulated oralsolutions. Formulations can be designed for immediate release orcontrolled release. Preferably, such oral formulations are not producedfrom a liquid form of the sodium salt of diazoxide.

Formulations of the salts of the compounds of Formulae I-VIII can alsobe prepared for transdermal, intranasal and intravenous (I.V.)administration, provided that when the anion is diazoxide and the cationis sodium, the formulation is not for intravenous use.

In another embodiment, formulations of the salts of the compounds ofFormulae I-VIII are prepared for transdermal or intranasal,administration, provided that when the anion is diazoxide and the cationis sodium, the formulation is not produced using a liquid form of thesalt of the compounds of Formulae I-VIII.

In another embodiment, formulations of the salts of the compounds ofFormulae I-VIII are prepared for transdermal, intranasal and intravenous(I.V.) administration excluding the sodium salt of diazoxide.

Formulations of K_(ATP) channel openers prepared using salts of thecompounds selected from Formulae I-VIII exhibit improved solubility andabsorption compared to previous formulations of these drugs. Theseadvantageous properties are achieved by any one or more of the followingapproaches: (1) reducing particle size of the formulation bycomminution, spray drying, or other micronising techniques, (2) using anion exchange resin in the formulation, (3) using inclusion complexes,for example using a cyclodextrin, (4) compaction of the salt of K_(ATP)channel opener with a solubilizing agent including low viscosityhypromellose, low viscosity methylcellulose or similarly functioningexcipient and combinations thereof, (5) associating the salt of theK_(ATP) channel opener with a distinct salt prior to formulation, (6)using a solid dispersion of the salt of the K_(ATP) channel opener, (7)using a self emulsifying system, (8) adding one or more surfactants tothe formulation, (9) using nanoparticles in the formulation, or (10)combinations of these approaches.

Release of K_(ATP) channel opener selected from salts of the compoundsof Formulae I-VIII over a sustained period of time (e.g., 2-30 hours)can be achieved by the use of one or more approaches including, but notlimited to: (1) the use of pH sensitive polymeric coatings, (2) the useof a hydrogel, (3) the use of a film coating that controls the rate ofdiffusion of the drug from a coated matrix, (4) the use of an erodablematrix that controls rate of drug release, (5) the use of polymer coatedpellets, granules, or microparticles which can be further encapsulatedor compressed into a tablet, (6) the use of an osmotic pump system, (7)the use of a compression coated tablet, or (8) combinations of theseapproaches.

Delay of release of K_(ATP) channel openers selected from the salts ofthe compounds of Formulae I-VIII from the formulation until gastrictransit is complete can be achieved in the formulations provided hereinby any of several mechanisms. For example, pH sensitive polymer orco-polymer can be used which when applied around the drug matrixfunctions as an effective barrier to release of active at pH 3.0 orlower and is unstable at pH 5.5 and above. This provides for control ofrelease of the active compound in the stomach but rapidly allows releaseonce the dosage form has passed into the small intestine. An alternativeto a pH sensitive polymer or co-polymer is a polymer or co-polymer thatis non-aqueous-soluble. The extent of resistance to release in thegastric environment can be controlled by coating with a blend of thenon-aqueous-soluble and a aqueous soluble polymer. In this approachneither of the blended polymers or co-polymers are pH sensitive. Oneexample of a pH sensitive co-polymer is the Eudragit® methacrylicco-polymers, including Eudragit® L 100, S 100 or L 100-55 solids, L 30D-55 or FS 30D dispersions, or the L 12.5 or S 12.5 organic solutions.

Polymers that delay release can be applied to a tablet either by spraycoating (as a thin film) or by compression coating. If a capsule isused, then the polymer(s) may be applied over the surface of the capsuleor applied to microparticles of the drug, which may then be encapsulatedsuch as in a capsule or gel. If the capsule is coated, then it willresist disintegration until after gastric transit. If microparticles arecoated, then the capsule may disintegrate in the stomach but little tono drug will be released until after the free microparticles completegastric transit. Finally, an osmotic pump system that uses e.g., aswellable hydrogel can be used to delay drug release in the stomach. Theswellable hydrogel takes up moisture after administration. Swelling ofthe gel results in displacement of the drug from the system forabsorption. The timing and rate of release of the drug depend on the gelused, and the rate at which moisture reaches the gel, which can becontrolled by the size of the opening in the system through which fluidenters. See Drug Delivery Technologies online article Dong et al.,“L-OROS® SOFTCAP™ for Controlled Release of Non-Aqueous LiquidFormulations.”

Accordingly, delay of release of formulations of K_(ATP) channel openersprepared as salts of the compounds of Formulae I-VIII until aftergastric transit is complete can be achieved by any of severalmechanisms, including, but not limited to: (a) a pH sensitive polymer orco-polymer applied as a compression coating on a tablet; (b) a pHsensitive polymer or co-polymer applied as a thin film on a tablet; (c)a pH sensitive polymer or co-polymer applied as a thin film to anencapsulation system; (d) a pH sensitive polymer or co-polymer appliedto encapsulated microparticles, (e) a non-aqueous-soluble polymer orcopolymer applied as a compression coating on a tablet; (f) anon-aqueous-soluble polymer or co-polymer applied as a thin film on atablet; (g) a non-aqueous soluble polymer applied as a thin film to anencapsulation system; (h) a non-aqueous soluble polymer applied tomicroparticles; (i) incorporation of the formulation in an osmotic pumpsystem, or (j) use of systems controlled by ion exchange resins, or (k)combinations of these approaches, wherein the pH sensitive polymer orco-polymer is resistant to degradation under acid conditions.

Formulations are provided that are designed for administration oncedaily (i.e., once per 24 hours). These formulations can contain between25 and 500 mg of K_(ATP) channel openers selected from salts of thecompounds of Formulae I-VIII. Formulations intended for administrationtwice daily (per 24 hours) may also be provided. These can containbetween 25 and 250 mg of K_(ATP) channel openers.

The formulations provided herein exhibit improved safety of theadministered drug product. This improvement in safety occurs by at leasttwo mechanisms. First, delay of release of active drug until gastrictransit is complete can reduce the incidence of a range ofgastrointestinal adverse side effects including nausea, vomiting,dyspepsia, abdominal pain, diarrhea and ileus. Second, by sustainingrelease of the active drug over 2 or more hours up to as long as 24hours, peak drug levels are reduced relative to the peak drug levelsobserved for the same administered dose using any oral formulation thatdoes not have sustained or controlled release. This reduction in peakdrug levels can contribute to reductions in adverse effects that arepartially or completely determined by peak drug levels. These adverseeffects include: fluid retention with the associated reduced rates ofexcretion of sodium, chloride and uric acid, edema, hyperglycemia andthe associated potential for progression to ketoacidosis, cataracts andnon-ketotic hyperosmolar coma, headaches, tachycardia and palpitations.

Also provided herein are controlled release formulations of K_(ATP)channel openers prepared from salts of compounds of Formulae I-VIII,which have one feature from each of A-D as shown in Table 1.

TABLE 1 Controlled Release Formulation Characteristics and Properties A.Unit Form: Tablet or Capsule B. Dosage/unit: 10-100 mg 100-200 mg200-300 mg 300-500 mg 500-2000 mg C. Dosing Once daily (24 hours) Twicedaily (24 hours) D. Release time: 2-4 hrs 4-8 hrs 8-24 hours

For example, a controlled release composition can be a tablet containing25-100 mg of a salt of a compound of Formulae I-VIII, wherein suchtablet administered once daily to achieve a controlled release time of2-4 hours. All of these formulations can further include the feature ofsubstantially delaying pharmaceutical active release until after gastrictransit is complete.

In addition, any of the above formulations from Table 1 can include atleast one feature that improves the solubility or absorption of theK_(ATP) channel opener.

Exemplary controlled release formulations provided herein include theactive compound (i.e., a K_(ATP) channel opener selected from a salt ofa compound of any of Formulae I-VIII) and a matrix which includes agelling agent that swells upon contact with aqueous fluid. The activecompound entrapped within the gel is slowly released into the body upondissolution of the gel. The active compound can be evenly dispersedwithin the matrix or can be present as pockets of drug in the matrix.For example, the drug can be formulated into small granules which aredispersed within the matrix. In addition, the granules of drug also caninclude a matrix, thus, providing a primary and a secondary matrix asdescribed in U.S. Pat. No. 4,880,830 to Rhodes.

The gelling agent preferably is a polymeric material, which can include,for example, any pharmaceutically acceptable water soluble or waterinsoluble slow releasing polymer such as xantham gum, gelatin, celluloseethers, gum arabic, locust bean gum, guar gum, carboxyvinyl polymer,agar, acacia gum, tragacanth, veegum, sodium alginate or alginic acid,polyvinylpyrrolidone, polyvinyl alcohol, or film forming polymers suchas methyl cellulose (MC), carboxymethyl cellulose (CMC), hydroxypropylmethylcellulose, hyroxypropyl methyl cellulose (HPMC), hydroxypropylcellulose (HPC), hydroxyethyl cellulose (HEC), ethylcellulose (EC),acrylic resins or mixtures of the above (see e.g., U.S. Pat. No.5,415,871).

The gelling agent of the matrix also can be a heterodisperse gumcomprising a heteropolysaccharide component and a homopolysaccharidecomponent which produces a fast-forming and rigid gel as described inU.S. Pat. No. 5,399,359. The matrix also can include a cross-linkingagent such as a monovalent or multivalent metal cations to further addrigidity and decrease dissolution of the matrix, thus further slowingrelease of drug. The amount of crosslinking agent to add can bedetermined using methods routine to the ordinary skilled artisan.

The matrix of the controlled release composition also can include one ormore pharmaceutically acceptable excipients recognized by those skilledin the art, i.e. formulation excipients. Such excipients include, forexample, binders: polyvinylpyrrolidone, gelatin, starch paste,microcrystalline cellulose; diluents (or fillers): starch, sucrose,dextrose, lactose, fructose, xylitol, sorbitol, sodium chloride,dextrins, calcium phosphate, calcium sulphate; and lubricants: stearicacid, magnesium stearate, calcium stearate, Precirol® and flow aids forexample talc or colloidal silicon dioxide.

The matrix of the controlled release composition can further include ahydrophobic material which slows the hydration of the gelling agentwithout disrupting the hydrophilic nature of the matrix, as described inU.S. Pat. No. 5,399,359. The hydrophobic polymer can include, forexample, alkylcellulose such as ethylcellulose, other hydrophobiccellulosic materials, polymers or copolymers derived from acrylic ormethacrylic acid esters, copolymers of acrylic and methacrylic acidesters, zein, waxes, shellac, hydrogenated vegetable oils, waxes andwaxy substances such as carnauba wax, spermaceti wax, candellila wax,cocoa butter, cetosteryl alcohol, beeswax, ceresin, paraffin, myristylalcohol, stearyl alcohol, cetylalcohol and stearic acid, and any otherpharmaceutically acceptable hydrophobic material known to those skilledin the art.

The amount of hydrophobic material incorporated into the controlledrelease composition is that which is effective to slow the hydration ofthe gelling agent without disrupting the hydrophilic matrix formed uponexposure to an environmental fluid. In certain preferred embodiments,the hydrophobic material is included in the matrix in an amount fromabout 1 to about 20 percent by weight and replaces a correspondingamount of the formulation excipient. A solvent for the hydrophobicmaterial may be an aqueous or organic solvent, or mixtures thereof.

Examples of commercially available alkylcelluloses are Aquacoat®(aqueous dispersion of ethylcellulose available from FMC) and Surelease®(aqueous dispersion of ethylcellulose available from Colorcon). Examplesof commercially available acrylic polymers suitable for use as thehydrophobic material include Eudragit® RS and RL (copolymers of acrylicand methacrylic acid esters having a low content (e.g., 1:20 or 1:40) ofquaternary ammonium compounds).

The controlled release composition also can be coated to retard accessof liquids to the active compound and/or retard release of the activecompound through the film-coating. The film-coating can providecharacteristics of gastroresistance and enterosolubility by resistingrapid dissolution of the composition in the digestive tract. Thefilm-coating generally represents about 5-15% by weight of thecontrolled release composition. Preferably, the core by weightrepresents about 90% of the composition with the remaining 10% providedby the coating. Such coating can be a film-coating as is well known inthe art and include gels, waxes, fats, emulsifiers, combination of fatsand emulsifiers, polymers, starch, and the like.

Polymers and co-polymers are useful as thin film coatings. Solutioncoatings and dispersion coatings can be used to coat the activecompound, either alone or combined with a matrix. The coating ispreferably applied to the drug or drug and matrix combination as a solidcore of material as is well known in the art.

A solution for coating can include polymers in both organic solvent andaqueous solvent systems, and typically further including one or morecompounds that act as a plasticizer. Polymers useful for coatingcompositions include, for example, methylcellulose (Methocel® A; DowChemical Co.), hydroxypropylmethylcellulose with a molecular weightbetween 1,000 and 4,000,000 (Methocel® E; Dow Chemical Co. orPharmacoat®; Shin Etsu), hydroxypropyl cellulose with a molecular weightbetween 2,000 and 2,000,000, ethyl cellulose, cellulose acetate,cellulose triacetate, cellulose acetate butyrate, cellulose acetatephthalate, cellulose acetate trimellitate (Eastman Kodak),carboxymethylethyl cellulose (Duodcel®), hydroxypropyl methylcellulosephthalate, ethylcellulose, methylcellulose and, in general, cellulosicderivatives, polymethacrylic acid-methacrylic acid copolymer (Type A 1:1Eudragit L100; Type B 1:2 Eudragit S100; and Type C 1:1 EudragitL100-55, aqueous dispersion 30% solids, Eudragit L30D), poly(meth)acrylester: poly(ethyl acrylate, methyl methacrylate 2:1), Eudragit NE30Daqueous dispersion 30% solids, polyaminomethacrylate Eudragit E100,poly(trimethylammonioethyl methacrylate chloride) ammoniomethacrylatecopolymer, Eudragit RL30D and Eudragit RS30D, carboxyvinyl polymers,polyvinylalcohols, glucans scleroglucans, mannans, and xanthans.

Aqueous polymeric dispersions include Eudragit L30D and RS/RL30D, andNE30D, AQUACOAT® brand ethyl cellulose, Surelease brand ethyl cellulose,EC brand N-10F ethyl cellulose, Aquateric brand cellulose acetatephthalate, Coateric brand Poly(vinyl acetate phthalate), and Aqacoatbrand hydroxypropyl methylcellulose acetate succinate. Most of thesedispersions are latex, pseudolatex powder or micronized powder mediums.

A plasticizing agent may be included in the coating to improve theelasticity and the stability of the polymer film and to prevent changesin the polymer permeability over prolonged storage. Such changes mayaffect the drug release rate. Suitable conventional plasticizing agentsinclude, for example, diethyl phthalate, glycerol triacetate, acetylatedmonoglycerides, acetyltributylcitrate, acetyltriethyl citrate, castoroil, citric acid esters, dibutyl phthalate, dibutyl sebacate,diethyloxalate, diethyl malate, diethylfumarate, diethylphthalate,diethylsuccinate, diethylmalonate, diethyltartarate, dimethylphthalate,glycerin, glycerol, glyceryl triacetate, glyceryltributyrate, mineraloil and lanolin alcohols, petrolatum and lanolin alcohols, phthalic acidesters, polyethylene glycols, propylene glycol, rape oil, sesame oil,triacetin, tributyl citrate, triethyl citrate, and triethyl acetylcitrate, or a mixture of any two or more of the foregoing. Plasticizerswhich can be used for aqueous coatings include, for example, propyleneglycol, polyethylene glycol (PEG 400), triacetin, polysorbate 80,triethyl citrate, and diethyl d-tartrate.

A coating solution comprising a mixture of hydroxypropylmethylcelluloseand aqueous ethylcellulose (e.g. Aquacoat brand) as the polymer anddibutyl sebacate as plasticizer can be used for coating microparticles.(Aquacoat is an aqueous polymeric dispersion of ethylcellulose andcontains sodium lauryl sulfate and cetyl alcohol). Preferably, theplasticizer represents about 1-2% of the composition.

In addition to the polymers, the coating layer can include an excipientto assist in formulation of the coating solution. Such excipients mayinclude a lubricant or a wetting agent. Suitable lubricants asexcipients for the film coating include, for example, talc, calciumstearate, colloidal silicon dioxide, glycerin, magnesium stearate,mineral oil, polyethylene glycol, and zinc stearate, aluminum stearateor a mixture of any two or more of the foregoing. Suitable wettingagents include, for example, sodium lauryl sulfate, acacia, benzalkoniumchloride, cetomacrogol emulsifying wax, cetostearyl alcohol, cetylalcohol, cholesterol, diethanolamine, docusate sodium, sodium stearate,emulsifying wax, glyceryl monostearate, hydroxypropyl cellulose, lanolinalcohols, lecithin, mineral oil, monoethanolamine, poloxamer,polyoxyethylene alkyl ethers, polyoxyethylene castor oil derivatives,polyoxyethylene sorbitan fatty acid esters, polyoxyethylene stearates,propylene glycol alginate, sorbitan esters, stearyl alcohol andtriethanolamine, or a mixture of any two or more of the foregoing.

The specified tablet or capsule formulations of Table 1 may includeco-formulation with an obesity treating drug (in addition to a K_(ATP)channel opener selected from a salt of a compound of Formulae I-VIII).Obesity treating drugs that may be used include, but are not limited to,sibutramine hydrochloride (5-30 mg/unit), orlistat (50-360 mg/unit),phentermine hydrochloride or resin complex (15 to 40 mg/unit),zonisamide (100 to 600 mg/unit), topiramate (64 to 400 mg/unit),naltrexone hydrochloride (50 to 600 mg/unit), rimonabant (5 to 20mg/unit), ADP356 (5 to 25 mg/unit), ATL962 (20 to 400 mg/unit), orAOD9604 (1 to 10 mg/unit). These formulations are preferably used oncedaily. For a twice daily dosing, the amount of K_(ATP) channel openerselected from a salt of a compound of Formulae I-VIII is one half theamount included in the once daily formulation and the co-formulatedobesity treating drug is half of the amount specified. Alternativeobesity treating drugs may include, but are not limited to: selectiveserotonin 2c receptor agonists, dopamine antagonists, cannabinoid-1receptor antagonists, leptin analogues, leptin transport and/or leptinreceptor promoters, neuropeptide Y and agouti-related peptideantagonists, proopiomelanocortin and cocaine and amphetamine regulatedtranscript promoters, melanocyte-stimulating hormone analogues,melanocortin-4 receptor agonists, and agents that affect insulinmetabolism/activity, which can include protein-tyrosine phosphatase-1Binhibitors, peroxisome proliferator activated receptor antagonists,short-acting bromocriptine (ergoset), somatostatin agonists(octreotide), and adiponectin, gastrointestinal-neural pathway agents,including those that increase cholecystokinin activity, increaseglucagon-like peptide-1 activity (e.g., extendin 4, liraglutide,dipeptidyl peptidase IV inhibitors), and increase protein YY3-36activity and those that decrease ghrelin activity, as well as amylinanalogues, agents that may increase resting metabolic rate (“selective”β-3 stimulators/agonist, uncoupling protein homologues, and thyroidreceptor agonists), melanin concentrating hormone antagonists,phytostanol analogues, amylase inhibitors, growth hormone fragments,synthetic analogues of dehydroepiandrosterone sulfate, antagonists ofadipocyte 11 Bhydroxysteroid dehydrogenase type 1 activity,corticotropin releasing hormone agonists, inhibitors of fatty acidsynthesis, carboxypeptidase inhibitors, indanones/indanols,aminosterols, and other gastrointestinal lipase inhibitors.

The specified tablet or capsule formulations of Table 1 may includeco-formulation with a diabetes treating drug (in addition to a K_(ATP)channel opener selected from a salt of a compound of Formulae I-VIII).Diabetes treating drugs that may be used include, but are not limitedto, acarbose (50 to 300 mg/unit), miglitol (25 to 300 mg/unit),metformin hydrochloride (300 to 2000 mg/unit), repaglinide (1-16mg/unit), nateglinide (200 to 400 mg/unit), rosiglitazone (5 to 50mg/unit), metaglidasen (100 to 400 mg/unit) or any drug that improvesinsulin sensitivity, or improves glucose utilization and uptake. Theseformulations are preferably used once daily. For a twice daily dosing,the amount of the K_(ATP) channel opener selected from a salt of acompound of Formulae I-VIII is half the amount included in the oncedaily formulation and the co-formulated diabetes treating drug is halfof the amount specified.

The specified tablet or capsule formulations of Table 1 may includeco-formulation with a cholesterol lowering drug. Cholesterol loweringdrugs that may be used include, but are not limited to, pravastatin,simvastatin, atorvastatin, fluvastatin, rosuvastatin, or lovastatin (allat 10 to 80 mg/unit). These formulations are preferably used once daily.For a twice daily dosing, the amount of K_(ATP) channel opener selectedfrom a salt of a compound of Formulae I-VIII is preferably 25 to 200mg/unit and the co-formulated cholesterol lowering drug is half of theamount specified.

The specified tablet or capsule formulations of Table 1 may includeco-formulation with a depression treating drug. Depression treatingdrugs that may be used include, but are not limited to, citalopramhydrobromide (10 to 80 mg/unit), escitalopram hydrobromide (5 to 40mg/unit), fluvoxamine maleate (25 to 300 mg/unit), paroxetinehydrochloride (12.5 to 75 mg/unit), fluoxetine hydrochloride (30 to 100mg/unit), setraline hydrochloride (25 to 200 mg/unit), amitriptylinehydrochloride (10 to 200 mg/unit), desipramine hydrochloride (10 to 300mg/unit), nortriptyline hydrochloride (10 to 150 mg/unit), duloxetinehydrochloride (20 to 210 mg/unit), venlafaxine hydrochloride (37.5 to150 mg/unit), phenelzine sulfate (10 to 30 mg/unit), bupropionhydrochloride (200 to 400 mg/unit), or mirtazapine (7.5 to 90 mg/unit).These formulations are preferably used once daily. For a twice dailydosing, the amount of K_(ATP) channel opener selected from a salt of acompound of Formulae I-VIII is preferably half the amount included inthe once daily formulation and the co-formulated depression treatingdrug is half of the amount specified.

The specified tablet or capsule formulations of Table 1 may includeco-formulation with a hypertension treating drug. Hypertension treatingdrugs that may be used include, but are not limited, to enalaprilmaleate (2.5 to 40 mg/unit), captopril (2.5 to 150 mg/unit), lisinopril(10 to 40 mg/unit), benzaepril hydrochloride (10 to 80 mg/unit),quinapril hydrochloride (10 to 80 mg/unit), peridopril erbumine (4 to 8mg/unit), ramipril (1.25 to 20 mg/unit), trandolapril (1 to 8 mg/unit),fosinopril sodium (10 to 80 mg/unit), moexipril hydrochloride (5 to 20mg/unit), losartan potassium (25 to 200 mg/unit), irbesartan (75 to 600mg/unit), valsartan (40 to 600 mg/unit), candesartan cilexetil (4 to 64mg/unit), olmesartan medoxamil (5 to 80 mg/unit), telmisartan (20 to 160mg/unit), eprosartan mesylate (75 to 600 mg/unit), atenolol (25 to 200mg/unit), propranolol hydrochloride (10 to 180 mg/unit), metoprololtartrate, succinate or fumarate (each at 25 to 400 mg/unit), nadolol (20to 160 mg/unit), betaxolol hydrochloride (10 to 40 mg/unit), acebutololhydrochloride (200 to 800 mg/unit), pindolol (5 to 20 mg/unit),bisoprolol fumarate (5 to 20 mg/unit), nifedipine (15 to 100 mg/unit),felodipine (2.5 to 20 mg/unit), amlodipine besylate (2.5 to 20 mg/unit),nicardipine (10 to 40 mg/unit), nisoldipine (10 to 80 mg/unit),terazosin hydrochloride (1 to 20 mg/unit), doxazosin mesylate (4 to 16mg/unit), prazosin hydrochloride (2.5 to 10 mg/unit), or alfuzosinhydrochloride (10 to 20 mg/unit). These formulations are preferably usedonce daily. For a twice daily dosing, the amount of K_(ATP) channelopener is preferably half the amount included in the once dailyformulation and the co-formulated hypertension treating drug is half ofthe amount specified.

The specified tablet or capsule formulations of Table 1 may includeco-formulation with a diuretic to treat edema. Diuretics that may beused include, but are not limited to amiloride hydrochloride (1 to 10mg/unit), spironolactone (10 to 100 mg/unit), triamterene (25 to 200mg/unit), bumetanide (0.5 to 4 mg/unit), furosemide (10 to 160 mg/unit),ethacrynic acid or ethacrynate sodium (each at 10 to 50 mg/unit),torsemide (5 to 100 mg/unit), chlorthalidone (10 to 200 mg/unit),indapamide (1 to 5 mg/unit), hydrochlorothiazide (10 to 100 mg/unit),chlorothiazide (50 to 500 mg/unit), bendroflumethiazide (5 to 25mg/unit), hydroflumethiazide (10 to 50 mg/unit), mythyclothiazide (1 to5 mg/unit), or polythiazide (1 to 10 mg/unit). These formulations arepreferably used once daily. For a twice daily dosing, the amount ofK_(ATP) channel opener selected from a salt of a compound of FormulaeI-VIII is preferably half the amount included in the once dailyformulation and the co-formulated diuretic is half of the amountspecified.

The specified tablet or capsule formulations of Table 1 may includeco-formulation with a drug to treat inflammation or pain. Drugs fortreating inflammation or pain that may be used include, but are notlimited to aspirin (100 to 1000 mg/unit), tramadol hydrochloride (25 to150 mg/unit), gabapentin (100 to 800 mg/unit), acetaminophen (100 to1000 mg/unit), carbamazepine (100 to 400 mg/unit), ibuprofen (100 to1600 mg/unit), ketoprofen (12 to 200 mg/unit), fenprofen sodium (100 to600 mg/unit), flurbiprofen sodium or flurbiprofen (both at 50 to 200mg/unit), or combinations of any of these with a steroid or aspirin.These formulations are preferably used once daily. For a twice dailydosing, the amount of K_(ATP) channel opener selected from a salt of acompound of Formulae I-VIII is preferably half the amount included inthe once daily formulation and the co-formulated drug to treatinflammation or pain is half of the amount specified.

The specified tablet or capsule formulations of Table 1 may includeco-formulation with a drug to treat obesity associated co-morbiditiesinclude those specified above for treating diabetes, cholesterol,depression, hypertension and edema, or drugs to treat atherosclerosis,osteoarthritis, disc herniation, degeneration of knees and hips, breast,endometrial, cervical, colon, leukemia and prostate cancers,hyperlipidemia, asthma/reactive airway disease, gallstones, GERD,obstructive sleep apnea, obesity hypoventilation syndrome, recurrentventral hernias, menstrual irregularity and infertility.

The specified tablet or capsule formulations of Table 1 may includeco-formulation with an anti-psychotic drug. The combination may be usedto treat the psychotic condition and to treat or prevent weight gain,dyslipidemia or impaired glucose tolerance in the treated subject. Drugsfor treating various psychotic conditions that may be used include, butare not limited to, lithium or a salt thereof (250 to 2500 mg/unit),carbamazepine or a salt thereof (50 to 1200 mg/unit), valproate,valproic acid, or divalproex (125 to 2500 mg/unit), lamotrigine (12.5 to200 mg/unit), olanzapine (5 to 20 mg/unit), clozapine (12.5 to 450mg/unit), or risperidone (0.25 to 4 mg/unit). These coformulations arepreferably intended for once per day administration. For a twice dailydosing, the amount of K_(ATP) channel opener selected from a salt of acompound of Formulae I-VIII is preferably half the amount included inthe once daily formulation and the co-formulated anti-psychotic is halfof the amount specified.

The specified tablet or capsule formulations of Table 1 may includeco-formulation with a drug to treat or prevent ischemic or reperfusioninjury. Drugs for treating or preventing ischemic or reperfusion injurythat may be used include, but are not limited to: low molecular weightheparins (e.g., dalteparin, enoxaparin, nadroparin, tinzaparin ordanaparoid), ancrod, pentoxifylline, nimodipine, flunarizine, ebselen,tirilazad, clomethiazole, an AMPA agonist (e.g., GYKI 52466, NBQX,YM90K, zonampanel, or MPQX), SYM 2081, selfotel, Cerestat, CP-101,606,dextrophan, dextromethorphan, MK-801, NPS 1502, remacemide, ACEA 1021,GV150526, eliprodil ifenprodil, lubeluzole, naloxone, nalmefeneciticoline, acetyl-1-carnitine, nifedipine, resveratrol, a nitronederivative, clopidogrel, dabigatram, prasugrel, troxoprodil, AGY-94806,or KAI-9803.

Provided are formulations administered once or twice daily to an obeseor overweight subject continuously result in a circulating concentrationof K_(ATP) channel opener selected from a salt of a compound of FormulaeI-VIII sufficient to induce weight loss. Weight loss occurs by thepreferential loss of body fat. Additional weight loss can occur when theformulation is administered in combination with a reduced calorie diet.

Provided are formulations of K_(ATP) channel openers selected from asalt of a compound of Formulae I-VIII administered as a single dose toan obese, overweight or obesity-prone subject that result in theinhibition of fasting or glucose stimulated insulin secretion for about24 hours or for about 18 hours.

Provided are formulations of K_(ATP) channel openers selected from asalt of a compound of Formulae I-VIII administered as a single dose toan obese, overweight or obesity-prone subject that result in theelevation of energy expenditure for about 24 hours or for about 18hours.

Provided are formulations of K_(ATP) channel openers selected from asalt of a compound of Formulae I-VIII administered as a single dose toan obese, overweight or obesity-prone subject that result in theelevation of beta oxidation of fat for about 24 hours or for about 18hours.

Provided are formulations of K_(ATP) channel openers selected from asalt of a compound of Formulae I-VIII administered as a single dose toan obese, overweight or obesity-prone hyperphagic subject that result inthe inhibition of hyperphagia for about 24 hours or for about 18 hours.

Provided are formulations suitable for continuous administration once ortwice daily (per 24 hours) to a subject, resulting in a circulatingconcentration of K_(ATP) channel openers selected from a salt of acompound of Formulae I-VIII sufficient to induce either beta-cell restor improved insulin sensitivity or both. Such beta-cell rest andimprovements in insulin sensitivity can contribute to effectivetreatment of type I diabetes, type II diabetes and prediabetes. Suchbeta-cell rest and improvements in insulin sensitivity can contribute toeffective restoration of normal glucose tolerance in type II diabeticand prediabetic subjects.

The various pharmaceutical K_(ATP) channel opener formulations selectedfrom a salt of a compound of Formulae I-VIII have a variety ofapplications, including, but not limited to: (1) treatment of obesity;(2) prevention of weight gain in subjects who are predisposed toobesity; (3) treatment of hyperinsulinemia or hyperinsulinism; (4)treatment of hypoglycemia; (5) treatment of hyperlipidemia, (6)treatment of type II diabetes, (7) preservation of pancreatic functionin type I diabetics; (8) treatment of metabolic syndrome (or syndromeX); (9) prevention of the transition from prediabetes to diabetes, (10)correction of the defects in insulin secretion and insulin sensitivitycontributing to prediabetes and type II diabetes, (11) treatment ofpolycystic ovary syndrome, (12) prevention of ischemic or reperfusioninjury, (13) treat weight gain, dyslipidemia, or impairment of glucosetolerance in subjects treated with antipsychotics drugs, (14) preventweight gain, dyslipidemia, or impairment of glucose tolerance insubjects treated with antipsychotics drugs and (15) treatment of anydisease where hyperlipidemia, hyperinsulinemia, hyperinsulinism,hyperlipidemia, hyperphagia or obesity are contributing factors to theseverity or progression of the disease, including but not limited to,Prader Willi Syndrome, Froelich's syndrome, Cohen syndrome, SummitSyndrome, Alstrom Syndrome, Borjesen Syndrome, Bardet-Biedl Syndrome, orhyperlipoproteinemia type I, II, III, and IV.

In one embodiment, a K_(ATP) channel opener selected from a salt of acompound of Formulae I-VIII is administered to an overweight or obesesubject as an oral dosage once per 24 hours to induce weight loss. Infurther embodiments, the subject (a) is not a type I diabetic, (b) isnot a type II diabetic, (c) is not experiencing chronic, recurrent ordrug-induced hypoglycemia, (d) does not have metabolic syndrome, or (e)is not experiencing malignant hypertension.

In one embodiment, a K_(ATP) channel opener selected from a salt of acompound of Formulae I-VIII is administered to an overweight or obesesubject as an oral dosage twice per 24 hours to induce weight loss. Thistreatment can be the sole treatment to induce weight loss. In furtherembodiments, the overweight or obese subject (a) does not have aninsulin secreting tumor, (b) is not suffering from Poly Cystic OvarySyndrome, (c) is not a type I diabetic, (d) is not a type II diabetic,(e) does not have metabolic syndrome, (f) is not experiencing chronicrecurrent or drug-induced hypoglycemia, (g) has not been treated forschizophrenia with haloperidol, or (h) is not experiencing malignanthypertension. In further embodiments, the overweight or obese adolescent(a) has not been diagnosed as being type I or type II diabetic, (b) isnot experiencing chronic, recurrent or drug-induced hypoglycemia, or (c)has not been diagnosed as having metabolic syndrome.

In another embodiment, a K_(ATP) channel opener selected from a salt ofa compound of Formulae I-VIII is administered to an overweight or obesesubject as an oral dosage form three times per 24 hours to induce weightloss. This treatment can be the sole treatment to induce weight loss. Infurther embodiments, the overweight or obese subject (a) does not havean insulin-secreting tumor, (b) is not suffering from Poly Cystic OvarySyndrome, (c) is not a type I diabetic, (d) is not a type II diabetic,(e) does not have metabolic syndrome, or (f) is not experiencingchronic, recurrent or drug-induced hypoglycemia.

In another embodiment, a K_(ATP) channel opener selected from a salt ofa compound of Formulae I-VIII is administered to an overweight or obeseadolescent as an oral dosage form three times per 24 hours to induceweight loss. This treatment can be the sole treatment to induce weightloss. In further embodiments, the overweight or obese adolescent is (a)is not a type I or type II diabetic, (b) is not experiencing chronic,recurrent or drug-induced hypoglycemia or (c) does not have metabolicsyndrome.

In another embodiment, a K_(ATP) channel opener selected from a salt ofa compound of Formulae I-VIII is administered as an oral dosage formthree times per 24 hours to induce weight loss to an overweight or obeseadult who (a) is not simultaneously receiving glucagon injections,triiodothyroxin or furosemide, (b) is not being treated forschizophrenia with haloperidol, or (c) is not experiencing malignanthypertension.

In another embodiment, a K_(ATP) channel opener selected from a salt ofa compound of Formulae I-VIII is administered to an overweight or obesesubject as an oral dosage form four times per 24 hours to induce weightloss.

In another embodiment, a K_(ATP) channel opener selected from a salt ofa compound of Formulae I-VIII is administered to an overweight or obesesubject as an oral dosage form administered from one, two, three or fourtimes per 24 hours to induce weight loss at a daily dose of 50 to 700mg. In a further embodiment, the overweight or obese subject (a) is nottype I diabetic, (b) is not type II diabetic, (c) is not sufferingchronic, recurrent or drug-induced hypoglycemia, or (d) does not havemetabolic syndrome.

In another embodiment, a K_(ATP) channel opener selected from a salt ofa compound of Formulae I-VIII is administered to an overweight or obesesubject as an oral dosage form administered from one, two, three or fourtimes per 24 hours to induce weight loss at a daily dose of 130 to 400mg. In a further embodiment, the overweight or obese subject (a) is nottype I diabetic, (b) is not type II diabetic, (c) is not sufferingchronic, recurrent or drug-induced hypoglycemia, or (d) does not havemetabolic syndrome.

In other embodiments, a K_(ATP) channel opener selected from a salt of acompound of Formulae I-VIII is administered to an overweight or obesityprone subject as an oral dosage form one, two, three or four times per24 hours to maintain a weight loss, as it is preferable to maintainweight in an obese subject once some weight loss has occurred when thealternative is to regain weight. In a further embodiment, theadministered daily dose of the K_(ATP) channel opener is 50 to 275 mg.

In other embodiments, a K_(ATP) channel opener selected from a salt of acompound of Formulae I-VIII is administered as an oral dosage form to anoverweight, obese, or obesity prone subject to (a) elevate energyexpenditure, (b) elevate beta oxidation of fat, or (c) reducecirculating triglyceride concentrations.

In other embodiments, an oral dosage of a K_(ATP) channel openerselected from a salt of a compound of Formulae I-VIII is administered toan subject in need thereof to induce the loss of 25%, 50%, or 75% ofinitial body fat.

In another embodiment, an oral dosage of a K_(ATP) channel openerselected from a salt of a compound of Formulae I-VIII is administered toan subject in need thereof to induce (a) the preferential loss of bodyfat or (b) the preferential loss of visceral body fat.

In additional embodiments, an oral dosage of a K_(ATP) channel openerselected from a salt of a compound of Formulae I-VIII is administeredone, two or three times per 24 hours at daily doses of 50 to 700 mg toan subject to (a) induce the loss of 25%, 50% or 75% of initial bodyfat, (b) induce the preferential loss of body fat, or (c) induce thepreferential loss of visceral fat.

In another embodiment, an oral dosage of a K_(ATP) channel openerselected from a salt of a compound of Formulae I-VIII is administered toan subject to induce the preferential loss of body fat and to inducereduction in circulating triglycerides.

In another embodiment, an oral dosage of a K_(ATP) channel openerselected from a salt of a compound of Formulae I-VIII is co-administeredwith sibutramine, orlistat, rimonabant, an appetite suppressant, ananti-depressant, an anti-epileptic, a diuretic, a drug that inducesweight loss by a mechanism that is distinct from a K_(ATP) channelopener, or a drug that lowers blood pressure, to induce weight lossand/or treat obesity associated co-morbidities in an overweight, obese,or obesity prone subject. In further embodiments, the overweight, obese,or obesity prone subject (a) is a type I diabetic, (b) is not a type IIdiabetic, (c) is not suffering from chronic, recurrent or drug-inducedhypoglycemia, or (d) does not have metabolic syndrome.

In another embodiment an oral dosage of a K_(ATP) channel openerselected from a salt of a compound of Formulae I-VIII is co-administeredwith an anti-depressant, a drug that lowers blood pressure, a drug thatlowers cholesterol, a drug that raises HDL, an anti-inflammatory that isnot a Cox-2 inhibitor, a drug that lowers circulating triglycerides, toan overweight, obese, or obesity prone subject to induce weight lossand/or treat obesity associated co-morbidities. In further embodiments,the overweight, obese, or obesity prone subject (a) is not a type Idiabetic, (b) is not a type II diabetic, (c) is not suffering fromchronic, recurrent or drug-induced hypoglycemia, or (d) does not havemetabolic syndrome.

In another embodiment, an oral dosage of a K_(ATP) channel openerselected from a salt of a compound of Formulae I-VIII is co-administeredwith a drug that lowers blood pressure, a drug that lowers cholesterol,a drug that raises HDL, an anti-inflammatory that is not a Cox-2inhibitor, a drug that lowers circulating triglycerides, to maintainweight and/or treat obesity associated co-morbidities in an overweight,obese, or obesity prone subject, as it is preferable to maintain weightin an obese subject once some weight loss has occurred when thealternative is to regain weight. In further embodiments, the overweight,obese, or obesity prone subject (a) is not a type I diabetic, (b) is nota type II diabetic, (c) is not suffering from chronic, recurrent ordrug-induced hypoglycemia, or (d) does not have metabolic syndrome.

In additional embodiments, an oral dosage form of a K_(ATP) channelopener selected from a salt of a compound of Formulae I-VIII is used toadminister a therapeutically effective dose of a K_(ATP) channel openerto an obese, overweight or obesity prone subject in need thereof totreat obesity, to (a) provide beta cell rest, (b) treat type I or typeII diabetes, or (c) prevent the occurrence of diabetes.

In additional embodiments, an oral dosage form of a K_(ATP) channelopener selected from a salt of a compound of Formulae I-VIII isco-administered with Phentermine or a derivative thereof to an obeseadult or adolescent to induce weight loss and/or treat obesity andobesity-associated co-morbidities. In further embodiments, a solid oraldosage form or tablet formulation of a K_(ATP) channel opener isco-administered with Phentermine or a derivative thereof to an obeseadult or adolescent to treat metabolic syndrome in a patient in needthereof.

In further embodiments, a pharmaceutically acceptable formulation of aK_(ATP) channel opener selected from a salt of a compound of FormulaeI-VIII at doses of 50 to 700 mg/day is co-administered with Phentermineor a derivative thereof at daily doses of 15 to 37.5 mg to an overweightor obese subject to induce weight loss, to treat metabolic syndrome, orto induce weight loss and treat obesity-associated co-morbidities.

In another embodiment, a quick dissolving formulation of a K_(ATP)channel opener selected from a salt of a compound of Formulae I-VIII isused to provide a therapeutically effective dose to a patient in needthereof.

In further embodiments, a K_(ATP) channel opener selected from a salt ofa compound of Formulae I-VIII is administered once per 24 hours at dosesof 50 mg to 700 mg to an overweight or obese subject.

In further embodiments, a K_(ATP) channel opener selected from a salt ofa compound of Formulae I-VIII is formulated as a tablet or capsule fororal administration. The tablet or capsule may be co-formulated withmetformin. In another embodiment, a K_(ATP) channel opener selected froma salt of a compound of Formulae I-VIII is formulated as an oralsuspension or solution, and the oral suspension or solution may befurther encapsulated in another embodiment.

In another embodiment, a pharmaceutical salt of a K_(ATP) channel openerselected from a salt of a compound of Formulae I-VIII is formulated as atablet or capsule for oral administration, or as an oral suspension oras an oral solution, or as an oral suspension or solution that isencapsulated.

In another embodiment a K_(ATP) channel opener selected from a salt of acompound of Formulae I-VIII is co-formulated with hydro-chlorothiazide,chlorothiazide, cyclothiazide, benzthiazide, metyclothiazide,bendro-flumethiazide, hydroflumethiazide, trichlormethiazide, orpolythiazide in a pharmaceutical formulation suitable for oraladministration.

Upon administration of formulations which include a salt of a compoundof Formulae I-VIII provided herein to humans or animals, some or all ofthe following effects are observed: (1) the production of lipoproteinlipase by adipocytes is reduced; (2) enhanced lipolysis by adipocytes;(3) expression of fatty acid synthase by adipocytes is reduced; (4)glyceraldehydes phosphate dehydrogenase activity of adipocytes isreduced; (5) little or no new triglycerides are synthesized and storedby adipocytes; (6) enhanced expression of β3 Adrenergic Receptor (β3AR)an improvement in the adrenergic function in adipocytes; (7) reducedglucose stimulated secretion of insulin by pancreatic B-cells; (8)decreased insulinemia; (9) enhanced blood glucose levels; (10) increasedexpression of Uncoupling Protein 1 in adipocytes; (11) enhancedthermogenesis in white and brown adipose tissue; (12) reduction ofplasma triglyceride concentration; (13) decrease in circulating leptinconcentrations; (14) up-regulation of insulin receptors; (15) enhancedglucose uptake; (16) reduced adipocyte hyperplasia; (17) reducedadipocyte hypertrophy; (18) reduced rates of conversion of preadipocytesto adipocytes; (19) reduced rates of hyperphagia; (20) increasedprotection of CNS, cardiac and other tissues from ischemic orreperfusion injury; (21) improved insulin sensitivity; (22) elevated CSFinsulin concentrations; (23) elevated circulating adiponectinconcentrations; (25) reduced circulating triglyceride concentrations;(26) enhancement of beta-cell rest.

Threshold concentrations of the current invention include thosecirculating concentrations of K_(ATP) channel openers resulting from theadministration of salts of compounds of Formulae I-VIII as an i.v.formulation, an immediate release oral formulation, a controlled releaseformulation, a transdermal formulation, or an intranasal formulation toan overweight or obese subject which results in (1) measurablesuppression of fasting insulin levels, (2) suppression of fastinginsulin levels by at least 20% from the baseline measurement in the samesubject prior to treatment with a K_(ATP) channel opener selected from asalt of a compound of Formulae I-VIII, (3) suppression of fastinginsulin levels by at least 30% from the baseline measurement in the samesubject prior to treatment with a K_(ATP) channel opener selected from asalt of a compound of Formulae I-VIII, (4) suppression of fastinginsulin levels by at least 40% from the baseline measurement in the samesubject prior to treatment with a K_(ATP) channel opener selected from asalt of a compound of Formulae I-VIII, (5) suppression of fastinginsulin levels by at least 50% from the baseline measurement in the samesubject prior to treatment with a K_(ATP) channel opener selected from asalt of a compound of Formulae I-VIII, (6) suppression of fastinginsulin levels by at least 60% from the baseline measurement in the samesubject prior to treatment with a K_(ATP) channel opener selected from asalt of a compound of Formulae I-VIII, (7) suppression of fastinginsulin levels by at least 70% from the baseline measurement in the samesubject prior to treatment with a K_(ATP) channel opener selected from asalt of a compound of Formulae I-VIII, (8) suppression of fastinginsulin levels by at least 80% from the baseline measurement in the samesubject prior to treatment with a K_(ATP) channel opener selected from asalt of a compound of Formulae I-VIII, (9) loss of weight, (10)elevation of resting energy expenditure, or (11) elevation of theoxidation of fat or fatty acids. Threshold effects of the currentinvention include those circulating concentrations of K_(ATP) channelopeners selected from salts of compounds of Formulae I-VIII resultingfrom the administration of an i.v. formulation of the drug, or animmediate release oral formulation of the drug, or a controlled releaseformulation of the drug, or a sustained release formulation, or atransdermal formulation, or an intranasal formulation of the drug to anobesity prone subject which result in (1) the loss of weight, and (2)the maintenance of weight. Threshold effects of the current inventioninclude those circulating concentrations of K_(ATP) channel openersselected from salts of compounds of Formulae I-VIII resulting from theadministration of an i.v. formulation of the drug, or an immediaterelease oral formulation of the drug, or a controlled releaseformulation of the drug, or a sustained release formulation, or atransdermal formulation, or an intranasal formulation of the drug to aprediabetic subject which result in prevention of the transition todiabetes. Threshold effects of the current invention include thosecirculating concentrations of K_(ATP) channel openers resulting from theadministration of salts of compounds of Formulae I-VIII as an i.v.formulation, or an immediate release oral formulation, or a controlledrelease formulation, or a sustained release formulation, or atransdermal formulation, or an intranasal formulation to a subject withtype 1 diabetes which result in beta cell rest.

The mode of action by which weight is maintained or lost resulting fromthe prolonged administration of K_(ATP) channel openers selected fromsalts of compounds of Formulae I-VIII to overweight, obese or obesityprone subjects as provided herein includes, but is not limited to, oneor more of (1) enhanced energy expenditure, (2) enhanced oxidation offat and fatty acids, (3) enhancement of lipolysis in adipose tissue, (4)enhanced glucose uptake by tissues and enhanced insulin sensitivity, and(5) improved beta adrenergic response. The mode of action by whichweight is maintained or lost resulting from the prolonged administrationof K_(ATP) channel openers selected from salts of compounds of FormulaeI-VIII to obese or obesity prone subjects as provided herein may alsoinclude the suppression of appetite.

Prolonged administration of pharmaceutical formulations of K_(ATP)channel openers selected from salts of compounds of Formulae I-VIII tooverweight or obese humans or animals results in substantial andsustained weight loss including some or all of the following effects:(1) preferential loss of body fat; (2) loss of greater than 25% ofinitial body fat mass; (3) loss of greater than 50% of initial body fatmass; (4) loss of greater than 75% of initial body fat mass; (5)significant increase in resting energy expenditure; (6) increase in theoxidation of fat and fatty acids; (7) reduction in blood pressure; (8)production of lipoprotein lipase by adipocytes is reduced; (9) enhancedlipolysis by adipocytes; (10) expression of fatty acid synthase byadipocytes is reduced; (11) glyceraldehydes phosphate dehydrogenaseactivity of adipocytes is reduced; (12) little or no new triglyceridesare synthesized and stored by adipocytes; (13) enhanced expression of β₃Adrenergic Receptor (β₃AR) and an improvement in the adrenergic functionin adipocytes; (14) reduced glucose stimulated secretion of insulin bypancreatic B-cells; (15) decreased insulinemia; (16) enhanced bloodglucose levels; (17) increased expression of Uncoupling Protein 1 inadipocytes; (18) enhanced thermogenesis in white and brown adiposetissue; (19) reduction of plasma triglyceride concentration; (20)decrease in circulating leptin concentrations; (21) up-regulation ofinsulin receptors; (22) enhanced glucose uptake; (23) reduced adipocytehyperplasia; (24) reduced adipocyte hypertrophy; (25) reduced rates ofconversion of preadipocytes to adipocytes; (26) reduced rates ofhyperphagia; (27) the sequential loss first of the metabolically mostactive adipose tissue (visceral), followed by the loss of lessmetabolically active adipose tissue; (28) elevation of circulatingadiponectin concentrations; (29) elevation of cerebrospinal fluidinsulin levels; (30) enhanced islet insulin mRNA and insulin content; or(31) enhanced metabolic efficiency of insulin.

Prolonged administration of formulations of K_(ATP) channel openersselected from salts of compounds of Formulae I-VIII to obesity pronehumans or animals, including subjects who have undergone various typesof bariatric surgery, results in sustained maintenance of weightincluding some or all of the following effects: (1) increased restingenergy expenditure; (2) increase in the oxidation of fat and fattyacids; (3) reduction in blood pressure; (4) production of lipoproteinlipase by adipocytes is reduced; (5) enhanced lipolysis by adipocytes;(6) expression of fatty acid synthase by adipocytes is reduced; (7)glyceraldehyde phosphate dehydrogenase activity of adipocytes isreduced; (8) little or no new triglycerides are synthesized and storedby adipocytes; (9) enhanced expression of β₃ Adrenergic Receptor (β₃AR)and improvement in the adrenergic function in adipocytes; (10) reducedglucose stimulated secretion of insulin by pancreatic B-cells; (11)decreased insulinemia; (12) enhanced blood glucose levels; (13)increased expression of Uncoupling Protein 1 in adipocytes; (14)enhanced thermogenesis in white and brown adipose tissue; (15) reductionof plasma triglyceride concentration; (16) decreased circulating leptinconcentration; (17) up-regulation of insulin receptors; (18) enhancedglucose uptake; (19) reduced adipocyte hyperplasia; (20) reducedadipocyte hypertrophy; (21) reduced rates of conversion of preadipocytesto adipocytes; (22) reduced rates of hyperphagia; (23) elevatedcirculating adiponectin concentration; (24) elevated cerebrospinal fluidinsulin levels; (25) enhanced islet insulin mRNA and insulin content; or(26) enhanced metabolic efficiency of insulin.

Immediate or prolonged administration of formulations of K_(ATP) channelopeners selected from salts of compounds of Formulae I-VIII toprediabetic or type I diabetic humans or animals results in theprevention of beta cell failure, improved glycemic control, andprevention of the transition from prediabetes to diabetes including someor all of the following effects: (1) increase in resting energyexpenditure; (2) increase in the oxidation of fat and fatty acids; (3)reduction in blood pressure; (4) production of lipoprotein lipase byadipocytes is reduced; (5) enhanced lipolysis by adipocytes; (6)expression of fatty acid synthase by adipocytes is reduced; (7)glyceraldehyde phosphate dehydrogenase activity of adipocytes isreduced; (8) little or no new triglycerides are synthesized and storedby adipocytes; (9) enhanced expression of β₃ Adrenergic Receptor (β₃AR)and an improvement in the adrenergic function in adipocytes; (10)reduced glucose stimulated secretion of insulin by pancreatic B-cells;(11) decreased insulinemia; (12) enhanced blood glucose levels; (13)increased expression of Uncoupling Protein 1 in adipocytes; (14)enhanced thermogenesis in white and brown adipose tissue; (15) reductionof plasma triglyceride concentration; (16) decreased circulating leptinconcentrations; (17) up-regulation of insulin receptors; (18) enhancedglucose uptake; (19) reduced adipocyte hyperplasia; (20) reducedadipocyte hypertrophy; (21) reduced rates of conversion of preadipocytesto adipocytes; (22) reduced rates of hyperphagia; (23) elevatedcirculating adiponectin concentrations; (24) elevated cerebrospinalfluid insulin levels; (25) enhanced islet insulin mRNA and insulincontent; or (26) enhanced metabolic efficiency of insulin.

Immediate or prolonged administration of formulations of K_(ATP) channelopeners selected from salts of compounds of Formulae I-VIII to humans oranimals that are at risk for myocardial infarct, or stroke, orundergoing surgical procedure that restores blood flow to heart or brainresults in improved therapeutic outcomes post-surgically, or followingthe occurrence of myocardial infarct or stroke by improving the survivalof tissue after blood flow is restored, reduced stunning of tissue, andaltering the nature of the inflammatory responses.

Pharmaceutical formulations as provided herein are designed to be usedin the treatment of obesity, hyperlipidemia, hypertension, weightmaintenance, type I diabetes, prediabetes, type II diabetes, metabolicsyndrome or any condition where weight loss, reduction in circulatingtriglycerides or beta cell rest contributes to therapeutic outcomesprovide for a range of critical changes in pharmacodynamic andpharmacokinetic responses to administered doses of K_(ATP) channelopeners selected from salts of compounds of Formulae I-VIII whichchanges include one or more of the following: (1) extending thepharmacodynamic effect of an administered dose to 24 hours or longer asmeasured by the suppression of insulin secretion; (2) providing forsubstantial uptake of the active pharmaceutical ingredient in the smallintestine; (3) providing for substantial uptake of the activepharmaceutical ingredient in the large intestine; (4) result in loweredCmaxversus current oral suspension or capsule products for the sameadministered dose of active pharmaceutical ingredient; (5) provide forcirculating concentrations of unbound active pharmaceutical ingredientabove threshold concentrations for 24 or more hours from a singleadministered dose; and (6) provide for more consistent drug absorptionby treated subjects as compared to existing capsule formulations.

Pharmaceutical co-formulations of the current invention designed totreat a range of conditions in humans and animals include thecombination of K_(ATP) channel openers selected from salts of compoundsof Formulae I-VIII with: (1) a diuretic, (2) a drug that lowers bloodpressure, (3) a drug that suppresses appetite, (4) a cannabinoidreceptor antagonist, (5) a drug that suppresses that action of gastriclipases, (6) any drug that is used to induce weight loss, (7) a drugthat lowers cholesterol, (8) a drug that lowers LDL bound cholesterol,(9) a drug that improves insulin sensitivity, (10) a drug that improvesglucose utilization or uptake, (11) a drug that reduces incidence ofatherosclerotic plaque, (12) a drug that reduces inflammation, (13) adrug that is antidepressant, (14) a drug that is an anti-epileptic, or(15) a drug that is an anti-psychotic.

Treatment of humans or animals using pharmaceutical formulations (whichinclude K_(ATP) channel openers selected from salts of compounds ofFormulae I-VIII) result in reduced incidence of adverse side effectsincluding but not limited to edema, fluid retention, reduced rates ofexcretion of sodium, chloride, and uric acid, hyperglycemia,ketoacidosis, nausea, vomiting, dyspepsia, ileus and headaches. Thesereductions in frequency of adverse side effects are achieved by: (1)initiating dosing of subjects at subtherapeutic doses and in a step wisemanner increasing the dose daily until the therapeutic dose is achievedwhere the number of days over which the step up in dose is effected is 2to 10, (2) use of the lowest effective dose to achieve the desiredtherapeutic effect, (3) use of a pharmaceutical formulation that delaysrelease of active until gastric transit is complete, (4) use of apharmaceutical formulation that results in lower circulating peak druglevels as compared to an immediate release oral suspension or capsuleformulation for the same administered dose, and (5) optimizing thetiming of administration of dose within the day and relative to meals.

Treatment of patients suffering from Prader Willi Syndrome, Froelich'sSyndrome, Cohen Syndrome, Summit Syndrome, Alstrom Syndrome, BorjesenSyndrome, Bardet-Biedl Syndrome, and hyperlipoproteinemia type I, II,III, and IV with the current invention using pharmaceutical formulationsof K_(ATP) channel openers selected from salts of compounds of FormulaeI-VIII result in some or all of the following therapeutic outcomes: (1)weight loss, (2) reduced rates of weight gain, (3) inhibition ofhyperphagia, (4) reduced incidence of impaired glucose tolerance,prediabetes or diabetes, (5) reduced incidence of congestive heartfailure, (6) reduced hypertension, and (7) reduced rates of all causemortality.

Treatment of prediabetic subjects using invention pharmaceuticalformulations of K_(ATP) channel openers selected from salts of compoundsof Formulae I-VIII result in some or all of the following therapeuticoutcomes: (1) weight loss, (2) restoration of normal glucose tolerance,(3) delayed rates of progression to diabetes, (4) reduced hypertension,and (5) reduced rates of all cause mortality.

Treatment of diabetic subjects using invention pharmaceuticalformulations of K_(ATP) channel openers selected from salts of compoundsof Formulae I-VIII result in some or all of the following therapeuticoutcomes: (1) weight loss, (2) restoration of normal glucose tolerance,(3) delayed rates of progression of diabetes, (4) improvements inglucose tolerance, (5) reduced hypertension, and (6) reduced rates ofall cause mortality.

Co-administration of drugs with formulations of K_(ATP) channel openersselected from salts of compounds of Formulae I-VIII in the treatment ofdiseases of overweight, obese or obesity prone human and animal subjectsinvolves the co-administration of a pharmaceutically acceptableformulation of K_(ATP) channel openers with an acceptable formulationof: (1) sibutramine, (2) orlistat, (3) rimonabant, (4) a drug that is anappetite suppressant, (5) any drug used to induce weight loss in anobese or overweight subject, (6) a non-thiazide diuretic, (7) a drugthat lowers cholesterol, (8) a drug that raises HDL cholesterol, (9) adrug that lowers LDL cholesterol, (10) a drug that lowers bloodpressure, (11) a drug that is an anti-depressant, (12) a drug thatimproves insulin sensitivity, (13) a drug that improves glucoseutilization and uptake (14) a drug that is an anti-epileptic, (15) adrug that is an anti-inflammatory, or (16) a drug that lowerscirculating triglycerides.

Co-administration of drugs with formulations of K_(ATP) channel openersselected from salts of compounds of Formulae I-VIII in the treatment orprevention of weight gain, dyslipidemia, impaired glucose tolerance ordiabetes in subjects treated with antipsychotics drugs involve theco-administration of a pharmaceutically acceptable formulation ofK_(ATP) channel openers with an acceptable formulation of: lithium,carbamazepine, valproic acid and divalproex, and lamotrigine;antidepressants generally classified as monoamine oxidase inhibitorsincluding isocarboxazid, phenelzine sulfate and tranylcypromine sulfate;tricyclic antidepressants including doxepin, clomipramine,amitriptyline, maprotiline, desipromine, nortryptyline, desipramine,doxepin, trimipramine, imipramine and protryptyline; tetracyclicantidepressants including mianserin, mirtazapine, maprotiline,oxaprotiline, delequamine, levoprotiline, triflucarbine, setiptiline,azipramine, aptazapine maleate and pirlindole; and major tranquilizersand atypical antipsychotics including perphenazine, thioridazine,risperidone, clozapine, olanzapine and chlorpromazine.

In one embodiment, a K_(ATP) channel opener selected from a salt of acompound of Formulae I-VIII is administered to an overweight or obesesubject as an oral, transdermal or intranasal formulation to reach andmaintain the threshold concentration required to measurably reducefasting insulin levels for a prolonged period. Preferably the K_(ATP)channel opener formulation reduces fasting insulin levels by at least20%, more preferably by at least 30%, more preferably by at least by40%, more preferably by at least 50%, more preferably by at least by60%, more preferably by at least by 70%, and more preferably by at least80%. Fasting insulin levels are commonly measured using the glucosetolerance test (OGTT). After an overnight fast, a patient ingests aknown amount of glucose. Initial glucose levels are determined bymeasuring pre-test glucose levels in blood and urine. Blood insulinlevels are measured by a blood drop draw every hour after the glucose isconsumed for up to three hours. In a fasting glucose assay, subjectswith plasma glucose values greater than 200 mg/dl at 2 hourspost-glucose load indicate an impaired glucose tolerance.

In another embodiment, a K_(ATP) channel opener selected from a salt ofa compound of Formulae I-VIII is administered to an overweight or obesesubject as an oral, transdermal or intranasal formulation to reach andmaintain the threshold concentration required to induce weight loss fora prolonged period.

In another embodiment, a K_(ATP) channel opener selected from a salt ofa compound of Formulae I-VIII is administered to an overweight or obesesubject as an oral, transdermal or intranasal formulation to reach andmaintain the threshold concentration required to elevate resting energyexpenditure for a prolonged period.

In another embodiment, a K_(ATP) channel opener selected from a salt ofa compound of Formulae I-VIII is administered to an overweight or obesesubject as an oral, transdermal or intranasal formulation to reach andmaintain the threshold concentration required to elevate fat and fattyacid oxidation for a prolonged period.

In another embodiment, a K_(ATP) channel opener selected from a salt ofa compound of Formulae I-VIII is administered to an obesity pronesubject as an oral, transdermal or intranasal formulation to reach andmaintain the threshold concentration required to induce weight loss fora prolonged period.

In another embodiment, a K_(ATP) channel opener selected from a salt ofa compound of Formulae I-VIII is administered to an obesity pronesubject as an oral, transdermal or intranasal formulation to reach andmaintain the threshold concentration required to maintain weight for aprolonged period.

In another embodiment, a K_(ATP) channel opener selected from a salt ofa compound of Formulae I-VIII is administered to an overweight or obesesubject as an oral, transdermal or intranasal formulation to reach andmaintain a drug concentration above the threshold concentration requiredto induce weight loss for a prolonged period.

In another embodiment, a K_(ATP) channel opener selected from a salt ofa compound of Formulae I-VIII is administered to an overweight or obesesubject as an oral, transdermal or intranasal formulation for aprolonged period of time to reduce body fat by more than 25%, morepreferably by at least 50%, and more preferably by at least 75%.

In another embodiment, a K_(ATP) channel opener selected from a salt ofa compound of Formulae I-VIII is administered to an overweight or obesesubject as an oral, transdermal or intranasal formulation for aprolonged period of time to preferentially reduce visceral fat deposits.

In another embodiment, a K_(ATP) channel opener selected from a salt ofa compound of Formulae I-VIII is administered to an overweight or obesesubject as an oral, transdermal or intranasal formulation for aprolonged period of time to reduce visceral fat depots and other fatdeposits.

In another embodiment, a K_(ATP) channel opener selected from a salt ofa compound of Formulae I-VIII is administered to a normoinsulinemicoverweight or obese subject as an oral, transdermal or intranasalformulation to reach and maintain a drug concentration above thethreshold concentration required to induce weight loss for a prolongedperiod.

In another embodiment, a K_(ATP) channel opener selected from a salt ofa compound of Formulae I-VIII is administered to a prediabetic subjectas an oral, transdermal or intranasal formulation to reach and maintaina drug concentration above the threshold concentration required toprevent the transition to diabetes for a prolonged period.

In another embodiment, a K_(ATP) channel opener selected from a salt ofa compound of Formulae I-VIII is administered to a type 1 diabeticsubject as an oral, transdermal or intranasal formulation to reach andmaintain a drug concentration above the threshold concentration requiredto induce beta cell rest for a prolonged period.

In another embodiment, a single dose of a pharmaceutically acceptableformulation of a K_(ATP) channel opener selected from a salt of acompound of Formulae I-VIII is administered to an subject in needthereof that results in circulating concentration of active drugsufficient to diminish the secretion of insulin for 24 or more hours.

In another embodiment, a pharmaceutically acceptable formulation of aK_(ATP) channel opener selected from a salt of a compound of FormulaeI-VIII is administered over a prolonged basis to an subject in needthereof no more than once per 24 hours that results in circulatingconcentration of active drug sufficient to diminish the secretion ofinsulin on a continuous basis.

In another embodiment, a single dose of a pharmaceutically acceptableformulation of a K_(ATP) channel opener selected from a salt of acompound of Formulae I-VIII is administered to an subject in needthereof that results in circulating concentration of active drugsufficient to elevate non-esterified fatty acids in circulation for 24or more hours.

In another embodiment, a pharmaceutically acceptable formulation of aK_(ATP) channel opener selected from a salt of a compound of FormulaeI-VIII is administered over a prolonged basis to an subject in needthereof no more than once per 24 hours that results in circulatingconcentration of active drug sufficient to elevate non-esterified fattyacids in circulation on a continuous basis.

In another embodiment, a single dose of a pharmaceutically acceptableformulation of a K_(ATP) channel opener selected from a salt of acompound of Formulae I-VIII is administered to an subject in needthereof that results in circulating concentration of active drugsufficient to treat hypoglycemia in circulation for 24 or more hours.

In another embodiment, a pharmaceutically acceptable formulation of aK_(ATP) channel opener selected from a salt of a compound of FormulaeI-VIII is administered over a prolonged basis to an subject in needthereof no more than once per 24 hours that results in circulatingconcentration of active drug sufficient to treat hypoglycemia on acontinuous basis.

In another embodiment, a pharmaceutically acceptable formulation of aK_(ATP) channel opener selected from a salt of a compound of FormulaeI-VIII is administered over a prolonged basis to an subject in needthereof no more than once per 24 hours that results in circulatingconcentration of active drug sufficient to induce weight loss on acontinuous basis.

In another embodiment, a pharmaceutically acceptable formulation of aK_(ATP) channel opener selected from a salt of a compound of FormulaeI-VIII is administered over a prolonged basis to an subject in needthereof no more than once per 24 hours that results in circulatingconcentration of active drug sufficient to maintain weight on acontinuous basis, as it is preferable to maintain weight in an obesesubject once some weight loss has occurred when the alternative is toregain weight.

In another embodiment, a pharmaceutically acceptable formulation of aK_(ATP) channel opener selected from a salt of a compound of FormulaeI-VIII is administered over a prolonged basis to an subject in needthereof no more than once per 24 hours that results in circulatingconcentration of active drug sufficient to reduce circulatingtriglyceride levels on a continuous basis.

In another embodiment, a single dose of a pharmaceutically acceptableformulation of a K_(ATP) channel opener selected from a salt of acompound of Formulae I-VIII is administered to an subject in needthereof that results in circulating concentration of active drugsufficient to reduce or prevent ischemic or reperfusion injury incirculation for 24 or more hours.

In another embodiment, a pharmaceutically acceptable formulation of aK_(ATP) channel opener selected from a salt of a compound of FormulaeI-VIII is administered over a prolonged basis to an subject in needthereof no more than once per 24 hours that results in circulatingconcentration of active drug sufficient reduce or prevent ischemic orreperfusion injury on a continuous basis.

In another embodiment, the frequency of adverse effects caused bytreatment with a K_(ATP) channel opener selected from a salt of acompound of Formulae I-VIII is reduced using a pharmaceuticallyacceptable formulation of diazoxide or its derivatives that isadministered to an subject in need thereof on a daily basis in which thefirst dose is known to be subtherapeutic and daily dose is subsequentlyincreased stepwise until the therapeutic dose is reached.

In another embodiment, the frequency of adverse effects caused bytreatment with a K_(ATP) channel opener selected from a salt of acompound of Formulae I-VIII is reduced using a pharmaceuticallyacceptable formulation that is administered to an subject in needthereof on a daily basis in which the active ingredient is not releasedfrom the formulation until gastric transit is complete.

In another embodiment, the frequency of adverse effects caused bytreatment with a K_(ATP) channel opener selected from a salt of acompound of Formulae I-VIII is reduced using a pharmaceuticallyacceptable formulation that is administered to an subject in needthereof on a daily basis in which the maximum circulating concentrationof active ingredient is lower than what would be realized by theadministration of the same dose using an oral suspension or capsuleformulation of Proglycem®.

In another embodiment, the frequency of adverse effects caused bytreatment with a K_(ATP) channel opener selected from a salt of acompound of Formulae I-VIII is reduced using a pharmaceuticallyacceptable formulation that is administered to an subject in needthereof on a daily basis in which the first dose is known to besubtherapeutic and daily dose is subsequently increased stepwise untilthe therapeutic dose is reached, the active ingredient is not releasefrom the formulation until gastric transit is complete and in which themaximum circulating concentration of active ingredient is lower thanwhat would be realized by the administration of the same dose using anoral suspension or capsule formulation of Proglycem®.

In another embodiment, the frequency of adverse effects caused bytreatment with a K_(ATP) channel opener selected from a salt of acompound of Formulae I-VIII is reduced using a pharmaceuticallyacceptable formulation that is administered to an overweight or obesesubject in need thereof on a daily basis in which the first dose isknown to be subtherapeutic and daily dose is subsequently increasedstepwise until the therapeutic dose is reached, the active ingredient isnot release from the formulation until gastric transit is complete, inwhich the maximum circulating concentration of active ingredient islower than what would be realized by the administration of the same doseusing an oral suspension or capsule formulation of Proglycem®, and inwhich the maximum dose is less than 5 mg/kg/day.

In another embodiment, the frequency of adverse effects caused bytreatment with a K_(ATP) channel opener selected from a salt of acompound of Formulae I-VIII is reduced using a pharmaceuticallyacceptable formulation that is administered to an overweight or obesesubject in need thereof on a daily basis in which the first dose isknown to be subtherapeutic and daily dose is subsequently increasedstepwise until the therapeutic dose is reached, the active ingredient isnot release from the formulation until gastric transit is complete, inwhich the maximum circulating concentration of active ingredient islower than what would be realized by the administration of the same doseusing an oral suspension or capsule formulation, and in which themaximum dose is less than 2.5 mg/kg/day.

In another embodiment, the treatment of an overweight or obese subjectis optimized for weight loss by administration of a pharmaceuticallyacceptable formulation of a K_(ATP) channel opener selected from a saltof a compound of Formulae I-VIII once per 24 hours in which the releaseof the active ingredient from the formulation has been modified toprovide continuous release for at least 6 hours.

In another embodiment, the treatment of an overweight or obese subjectis optimized for weight loss by administration of a pharmaceuticallyacceptable formulation of a K_(ATP) channel opener selected from a saltof a compound of Formulae I-VIII once per 24 hours in which the releaseof the active ingredient from the formulation has been modified toprovide continuous release for at least 12 hours.

In another embodiment, the treatment of an overweight or obese subjectis optimized for weight loss by administration of a pharmaceuticallyacceptable formulation of a K_(ATP) channel opener selected from a saltof a compound of Formulae I-VIII once per 24 hours in which the releaseof the active ingredient from the formulation has been modified toprovide a rising drug concentration in circulation for at least 8 hours.

In another embodiment, the treatment of an overweight or obese subjectis optimized for weight loss by administration of a pharmaceuticallyacceptable formulation of a K_(ATP) channel opener selected from a saltof a compound of Formulae I-VIII once per 24 hours in which the releaseof the active ingredient from the formulation has been modified toprovide a rising drug concentration in circulation for at least 12hours.

In another embodiment, the treatment of an overweight or obese subjectis optimized for weight loss by administration of a pharmaceuticallyacceptable formulation of a K_(ATP) channel opener selected from a saltof a compound of Formulae I-VIII once per 24 hours in which the releaseof the active ingredient from the formulation has been modified to matchthe pattern of basal insulin secretion.

In another embodiment, the frequency of adverse effects caused bytreatment with a K_(ATP) channel opener selected from a salt of acompound of Formulae I-VIII is reduced using a pharmaceuticallyacceptable formulation that is administered to an obesity prone subjectin need thereof on a daily basis in which the first dose is known to besubtherapeutic and daily dose is subsequently increased stepwise untilthe therapeutic dose is reached, the active ingredient is not releasefrom the formulation until gastric transit is complete, in which themaximum circulating concentration of active ingredient is lower thanwhat would be realized by the administration of the same dose using anoral suspension or capsule formulation, and in which the maximum dose isless than 5 mg/kg/day.

In another embodiment, the frequency of adverse effects caused bytreatment with a K_(ATP) channel opener selected from a salt of acompound of Formulae I-VIII is reduced using a pharmaceuticallyacceptable formulation that is administered to an obesity prone subjectin need thereof on a daily basis in which the first dose is known to besubtherapeutic and daily dose is subsequently increased stepwise untilthe therapeutic dose is reached, the active ingredient is not releasefrom the formulation until gastric transit is complete, in which themaximum circulating concentration of active ingredient is lower thanwhat would be realized by the administration of the same dose using anoral suspension or capsule formulation, and in which the maximum dose isless than 2.5 mg/kg/day.

In another embodiment, the treatment of an obesity prone subject isoptimized for weight maintenance by administration of a pharmaceuticallyacceptable formulation of a K_(ATP) channel opener selected from a saltof a compound of Formulae I-VIII once per 24 hours in which the releaseof the active ingredient from the formulation has been modified toprovide continuous release for at least 6 hours.

In another embodiment, the treatment of an obesity prone subject isoptimized for weight maintenance by administration of a pharmaceuticallyacceptable formulation of a K_(ATP) channel opener selected from a saltof a compound of Formulae I-VIII once per 24 hours in which the releaseof the active ingredient from the formulation has been modified toprovide continuous release for at least 12 hours.

In another embodiment, the treatment of an obesity prone subject isoptimized for weight maintenance by administration of a pharmaceuticallyacceptable formulation of a K_(ATP) channel opener selected from a saltof a compound of Formulae I-VIII once per 24 hours in which the releaseof the active ingredient from the formulation has been modified toprovide a rising drug concentration in circulation for at least 8 hours.

In another embodiment, the treatment of an obesity prone subject isoptimized for weight maintenance by administration of a pharmaceuticallyacceptable formulation of a K_(ATP) channel opener selected from a saltof a compound of Formulae I-VIII once per 24 hours in which the releaseof the active ingredient from the formulation has been modified toprovide a rising drug concentration in circulation for at least 12hours.

In another embodiment, the treatment of an obesity prone subject isoptimized for weight maintenance by administration of a pharmaceuticallyacceptable formulation of a K_(ATP) channel opener selected from a saltof a compound of Formulae I-VIII once per 24 hours in which the releaseof the active ingredient from the formulation has been modified to matchthe pattern of basal insulin secretion.

In another embodiment, a pharmaceutically acceptable formulation of aK_(ATP) channel opener selected from a salt of a compound of FormulaeI-VIII is co-administered with sibutramine to an overweight or obesesubject to induce weight loss.

In another embodiment, a pharmaceutically acceptable formulation of aK_(ATP) channel opener selected from a salt of a compound of FormulaeI-VIII is co-administered with orlistat to an overweight or obesesubject to induce weight loss.

In another embodiment, a pharmaceutically acceptable formulation of aK_(ATP) channel opener selected from a salt of a compound of FormulaeI-VIII is co-administered with rimonabant to an overweight or obesesubject to induce weight loss.

In another embodiment, a pharmaceutically acceptable formulation of aK_(ATP) channel opener selected from a salt of a compound of FormulaeI-VIII is co-administered with an appetite suppressant to an overweightor obese subject to induce weight loss.

In another embodiment, a pharmaceutically acceptable formulation of aK_(ATP) channel opener selected from a salt of a compound of FormulaeI-VIII is co-administered with an anti-depressant to an overweight orobese subject to induce weight loss.

In another embodiment, a pharmaceutically acceptable formulation of aK_(ATP) channel opener selected from a salt of a compound of FormulaeI-VIII is co-administered with anti-epileptic to an overweight or obesesubject to induce weight loss.

In another embodiment, a pharmaceutically acceptable formulation of aK_(ATP) channel opener selected from a salt of a compound of FormulaeI-VIII is co-administered with a non-thiazide diuretic to an overweightor obese subject to induce weight loss.

In another embodiment, a pharmaceutically acceptable formulation of aK_(ATP) channel opener selected from a salt of a compound of FormulaeI-VIII is co-administered with a drug that induces weight loss by amechanism that is distinct from diazoxide to an overweight or obesesubject to induce weight loss.

In another embodiment, a pharmaceutically acceptable formulation of aK_(ATP) channel opener selected from a salt of a compound of FormulaeI-VIII is co-administered with a drug that lowers blood pressure to anoverweight, obesity prone or obese subject to induce weight loss andtreat obesity associated co-morbidities.

In another embodiment, a pharmaceutically acceptable formulation of aK_(ATP) channel opener selected from a salt of a compound of FormulaeI-VIII is co-administered with a drug that lowers cholesterol to anoverweight, obesity prone or obese subject to induce weight loss andtreat obesity associated co-morbidities.

In another embodiment, a pharmaceutically acceptable formulation of aK_(ATP) channel opener selected from a salt of a compound of FormulaeI-VIII is co-administered with a drug that raises HDL associatedcholesterol to an overweight, obesity prone or obese subject to induceweight loss and treat obesity associated co-morbidities.

In another embodiment, a pharmaceutically acceptable formulation of aK_(ATP) channel opener selected from a salt of a compound of FormulaeI-VIII is co-administered with a drug that improves insulin sensitivityto an overweight, obesity prone or obese subject to induce weight lossand treat obesity associated co-morbidities.

In another embodiment, a pharmaceutically acceptable formulation of aK_(ATP) channel opener selected from a salt of a compound of FormulaeI-VIII is co-administered with a an anti-inflammatory to an overweight,obesity prone or obese subject to induce weight loss and treat obesityassociated co-morbidities.

In another embodiment, a pharmaceutically acceptable formulation of aK_(ATP) channel opener selected from a salt of a compound of FormulaeI-VIII is co-administered with a drug that lowers circulatingtriglycerides to an overweight, obesity prone or obese subject to induceweight loss and treat obesity associated co-morbidities.

In another embodiment, K_(ATP) channel openers selected from salts ofcompounds of Formulae I-VIII are co-formulated with sibutramine in apharmaceutically acceptable formulation that is administered to anoverweight, obesity prone or obese subject to induce weight loss andtreat obesity-associated co-morbidities.

In another embodiment, K_(ATP) channel openers selected from salts ofcompounds of Formulae I-VIII are co-formulated with orlistat or otheractive that suppresses the action of gastric lipases in apharmaceutically acceptable formulation that is administered to anoverweight, obesity prone or obese subject to induce weight loss andtreat obesity-associated co-morbidities.

In another embodiment, K_(ATP) channel openers selected from salts ofcompounds of Formulae I-VIII are co-formulated with a non-thiazidediuretic in a pharmaceutically acceptable formulation that isadministered to an overweight, obesity prone or obese subject to induceweight loss and treat obesity-associated co-morbidities.

In another embodiment, K_(ATP) channel openers selected from salts ofcompounds of Formulae I-VIII are co-formulated with an appetitesuppressant in a pharmaceutically acceptable formulation that isadministered to an overweight, obesity prone or obese subject to induceweight loss and treat obesity-associated co-morbidities.

In another embodiment, K_(ATP) channel openers selected from salts ofcompounds of Formulae I-VIII are co-formulated with a cannabinoidreceptor antagonist in a pharmaceutically acceptable formulation that isadministered to an overweight, obesity prone or obese subject to induceweight loss and treat obesity-associated co-morbidities.

In another embodiment, K_(ATP) channel openers selected from salts ofcompounds of Formulae I-VIII are co-formulated with ananti-cholesteremic active in a pharmaceutically acceptable formulationthat is administered to an overweight, obesity prone or obese subject toinduce weight loss and treat obesity-associated co-morbidities.

In another embodiment, K_(ATP) channel openers selected from salts ofcompounds of Formulae I-VIII are co-formulated with an antihypertensiveactive in a pharmaceutically acceptable formulation that is administeredto an overweight, obesity prone or obese subject to induce weight lossand treat obesity-associated co-morbidities.

In another embodiment, K_(ATP) channel openers selected from salts ofcompounds of Formulae I-VIII are co-formulated with an insulinsensitizing active in a pharmaceutically acceptable formulation that isadministered to an overweight, obesity prone or obese subject to induceweight loss and treat obesity-associated co-morbidities.

In another embodiment, K_(ATP) channel openers selected from salts ofcompounds of Formulae I-VIII are co-formulated with an anti-inflammatoryactive in a pharmaceutically acceptable formulation that is administeredto an overweight, obesity prone or obese subject to induce weight lossand treat obesity-associated co-morbidities.

In another embodiment, K_(ATP) channel openers selected from salts ofcompounds of Formulae I-VIII are co-formulated with an anti-depressantactive in a pharmaceutically acceptable formulation that is administeredto an overweight, obesity prone or obese subject to induce weight lossand treat obesity-associated co-morbidities.

In another embodiment, K_(ATP) channel openers selected from salts ofcompounds of Formulae I-VIII are co-formulated with an anti-epilepticactive in a pharmaceutically acceptable formulation that is administeredto an overweight, obesity prone or obese subject to induce weight lossand treat obesity-associated co-morbidities.

In another embodiment, K_(ATP) channel openers selected from salts ofcompounds of Formulae I-VIII are co-formulated with an active thatreduces the incidence of atherosclerotic plaque in a pharmaceuticallyacceptable formulation that is administered to an overweight, obesityprone or obese subject to induce weight loss and treatobesity-associated co-morbidities.

In another embodiment, K_(ATP) channel openers selected from salts ofcompounds of Formulae I-VIII are co-formulated with an active thatlowers circulating concentrations of triglycerides in a pharmaceuticallyacceptable formulation that is administered to an overweight, obesityprone or obese subject to induce weight loss and treatobesity-associated co-morbidities.

The reduction of circulating triglycerides in an overweight, obese orobesity prone subject is achieved by the administration of an effectiveamount of an oral dosage form of a K_(ATP) channel opener selected froma salt of a compound of Formulae I-VIII.

An oral dosage form of K_(ATP) channel opener selected from a salt of acompound of Formulae I-VIII can be used to administer a therapeuticallyeffective dose of K_(ATP) channel opener to an overweight or obesityprone subject in need thereof to maintain weight, as it is preferable tomaintain weight in an obese subject once some weight loss has occurredwhen the alternative is to regain weight.

In another embodiment of the invention, K_(ATP) channel openers selectedfrom salts of compounds of Formulae I-VIII are co-formulated with a drugto treat obesity. Such co-formulations can be formulated for oraladministration once per 24 hours, for delayed release of the activeuntil gastric transit is complete, and for sustained release of theactive over a period of 2 to 24 hours. Such obesity treatment drugsinclude, but are not limited to: sibutramine hydrochloride (5-30 mg),orlistat (50-360 mg), phentermine hydrochloride or resin complex (15 to40 mg), zonisamide (100 to 600 mg), topiramate (64 to 400 mg),naltrexone hydrochloride (50 to 600 mg), or rimonabant (5 to 20 mg).

A further embodiment of the co-formulation contains K_(ATP) channelopeners selected from salts of compounds of Formulae I-VIII and a drugto treat obesity. Such co-formulations can be formulated for oraladministration twice per 24 hours, for delayed release of the activeuntil gastric transit is complete, and for sustained release of theactive over a period of 2 to 12 hours. Such obesity treatment drugsinclude, but are not limited to: sibutramine hydrochloride (2.5 to 15mg), orlistat (25 to 180 mg), phentermine hydrochloride or resin complex(7.5 to 20 mg), zonisamide (50 to 300 mg), topiramate (32 to 200 mg),naltrexone hydrochloride (25 to 300 mg), or rimonabant (2.5 to 10 mg).

In another embodiment of the invention K_(ATP) channel openers selectedfrom salts of compounds of Formulas I-VIII are co-formulated with a drugto treat obesity, diabetes, metabolic syndrome or an obesity relatedcomorbidity. Such drugs to treat these conditions include drugs that:agonizes the α1-noradrenergic receptor; agonizes the β2 noradrenergicreceptor; stimulates noradrenalin release; blocks noradrenalin uptake;stimulates 5-HT release; blocks 5-HT uptake; is a serotonin(5-hydroxytryptamine) 2C receptor agonist; antagonizes acetyl-CoAcarboxylase 2; agonizes the D1-receptor; antagonizes the H3-receptor; isa leptin analogue; agonizes the leptin receptor, sensitizes CNS tissueto the action of leptin; agonizes the MC4 receptor; agonizes NPY-Y1;agonizes NPY-Y2; agonizes NPY-Y4; agonizes NPY-Y5; antagonizes the MCHreceptor; blocks CRH-BP; agonizes the CRH receptor; agonizes theurocortin receptor; antagonizes the galanin receptor; antagonizes theorexin receptor; agonizes the CART receptor; agonizes the Amylinreceptor; agonizes the Apo(A^(IV)) receptor; antagonizes the CB-1receptor; is an aMSH analogue; inhibits PTP-1B; antagonizes PPARyreceptor; is a short acting bromocriptine; agonizes somatostatin;increases adiponectin; increases CCK activity; increases PYY activity;increases GLP-1 activity; decreases ghrelin activity; is a selective B3stimulator or agonist; agonizes thyroid receptor; inhibitsgastrointestinal lipases or other digestive enzymes; blocks absorptionof dietary fat; or block de-novo fatty acid synthesis. Additionally,such drugs to treat obesity may include, but are not limited to thosethat antagonize or agonize the function or expression of 11B□hydroxysteroid dehydrogenase type 1; acetyl-CoA carboxylase 1; ADAM 12,member 12 of a disintegrin and metalloprotease family or its shortersecreted form; agouti related protein; angiotensinogen; adipocyte lipidbinding protein; adipocyte fatty acid binding protein; adrenergicreceptors; acylation-stimulating protein; bombesin receptor subtype-3;C/EBP, CCAAT/enhancer binding protein; cocaine- andamphetamine-regulated transcript; cholecystokinin; cholecystokinin Areceptor; CD36, fatty acid translocase; corticotropin-releasing hormone;diacylglycerol acyltransferases; E2F transcription factor; eukaryotictranslation initiation factor 4e binding protein 1; estrogen receptor;fatty acid synthase; fibroblast growth factor; forkhead box C2;glucose-dependent insulinotropic peptide; GIP receptor; inhibitory Gprotein alpha-subunit; glucagon-like peptide-1; GLP-1 receptor;glycerol-3-phosphate acyltransferase; glycerol 3-phosphatedehydrogenase; stimulatory G protein alpha-subunit; high-mobility groupphosphoprotein isoform I-C; hormone sensitive lipase; inducible nitricoxide synthase; Janus kinases; lipoprotein lipase; melanocortin-3receptor; melanocortin-4 receptor; mitochondrial GPAT; metallothionein-Iand -II; nescient helix-loop-helix 2; neuropeptide Y; neuropeptide Y-1receptor; neuropeptide Y-2 receptor; neuropeptide Y-4 receptor;neuropeptide Y-5 receptor; plasminogen activator inhibitor-1;PPARgamma-co-activator 1; pro-opiomelanocortin; peroxisomeproliferator-activated receptor; protein tyrosine phosphatase 1B;regulatory subunit IIbeta □of protein kinase A; retinoid X receptor;steroidogenic factor 1; single-minded 1; sterol regulatory elementbinding protein; tyrosine hydroxylase; thyroid hormone receptor a2;uncoupling protein; nerve growth factor induced protein; leucine zippertranscription factor; a-melanocyte-stimulating hormone.

In another embodiment of the invention, K_(ATP) channel openers selectedfrom salts of compounds of Formulae I-VIII are co-formulated with a drugto treat diabetes. Such co-formulations can be formulated for oraladministration once per 24 hours, for delayed release of the activeuntil gastric transit is complete, and for sustained release of theactive over a period of 2 to 24 hours. Such diabetes treatment drugsinclude, but are not limited to: acarbose (50 to 300 mg), miglitol (25to 300 mg), metformin hydrochloride (300 to 2000 mg), repaglinide (1-16mg), nateglinide (200 to 400 mg), or rosiglitazone (5 to 50 mg).

In a further embodiment, the co-formulation can be formulated for oraladministration twice per 24 hours, for delayed release of the activeuntil gastric transit is complete, and for sustained release of theactive over a period of 2 to 12 hours. Such drugs to treat diabetesinclude, but are not limited to: acarbose (25 to 150 mg), miglitol (12.5to 150 mg), metformin hydrochloride (150 to 1000 mg), repaglinide (0.5to 8 mg), nateglinide (100 to 200 mg), or rosiglitizone (2.5 to 25 mg).

In another embodiment of the invention, K_(ATP) channel openers selectedfrom salts of compounds of Formulae I-VIII are co-formulated with a drugto treat elevated cholesterol. Such co-formulations can be formulatedfor oral administration once per 24 hours, for delayed release of theactive until gastric transit is complete, and for sustained release ofthe active over a period of 2 to 24 hours. Such drugs to treat elevatedcholesterol include, but are not limited to: pravastatin, simvastatin,atorvastatin, fluvastatin, rosuvastatin or lovastatin (10 to 80 mg).

In a further embodiment, the co-formulation can be formulated for oraladministration twice per 24 hours, for delayed release of the activeuntil gastric transit is complete, and for sustained release of theactive over a period of 2 to 12 hours. Such drugs to treat elevatedcholesterol include, but are not limited to: pravastatin, simvastatin,atorvastatin, fluvastatin, rosuvastatin or lovastatin (5 to 40 mg).

In another embodiment of the invention, K_(ATP) channel openers selectedfrom salts of compounds of Formulae I-VIII are co-formulated with a drugto treat depression. Such co-formulations can be formulated for oraladministration once per 24 hours, for delayed release of the activeuntil gastric transit is complete, and for sustained release of theactive over a period of 2 to 24 hours. Such drugs to treat depressioninclude, but are not limited to: citalopram hydrobromide (10 to 80 mg),escitalopram hydrobromide (5 to 40 mg), fluvoxamine maleate (25 to 300mg), paroxetine hydrochloride (12.5 to 75 mg), fluoxetine hydrochloride(30 to 100 mg), setraline hydrochloride (25 to 200 mg), amitriptylinehydrochloride (10 to 200 mg), desipramine hydrochloride (10 to 300 mg),nortriptyline hydrochloride (10 to 150 mg), duloxetine hydrochloride (20to 210 mg), venlafaxine hydrochloride (37.5 to 150 mg), phenelzinesulfate (10 to 30 mg), bupropion hydrochloride (200 to 400 mg), ormirtazapine (7.5 to 90 mg).

In a further embodiment, the co-formulation can be formulated for oraladministration twice per 24 hours, for delayed release of the activeuntil gastric transit is complete, and for sustained release of theactive for 2 to 12 hours. Such drugs to treat depression include, butare not limited to: citalopram hydrobromide (5 to 40 mg), escitalopramhydrobromide (2.5 to 20 mg), fluvoxamine maleate (12.5 to 150 mg),paroxetine hydrochloride (6.25 to 37.5 mg), fluoxetine hydrochloride (15to 50 mg), setraline hydrochloride (12.5 to 100 mg), amitriptylinehydrochloride (5 to 100 mg), desipramine hydrochloride (5 to 150 mg),nortriptyline hydrochloride (5 to 75 mg), duloxetine hydrochloride (10to 100 mg), venlafaxine hydrochloride (18 to 75 mg), phenelzine sulfate(5 to 15 mg), bupropion hydrochloride (100 to 200 mg), or mirtazapine (4to 45 mg).

In another embodiment of the invention, K_(ATP) channel openers selectedfrom salts of compounds of Formulae I-VIII are co-formulated with a drugto treat hypertension. Such co-formulations can be formulated for oraladministration once per 24 hours, for delayed release of the activeuntil gastric transit is complete, and for sustained release of theactive over a period of 2 to 24 hours. Such drugs to treat hypertensioninclude, but are not limited to: enalapril maleate (2.5 to 40 mg),captopril (2.5 to 150 mg), lisinopril (10 to 40 mg), benzaeprilhydrochloride (10 to 80 mg), quinapril hydrochloride (10 to 80 mg),peridopril erbumine (4 to 8 mg), ramipril (1.25 to 20 mg), trandolapril(1 to 8 mg), fosinopril sodium (10 to 80 mg), moexipril hydrochloride (5to 20 mg), losartan potassium (25 to 200 mg), irbesartan (75 to 600 mg),valsartan (40 to 600 mg), candesartan cilexetil (4 to 64 mg), olmesartanmedoxamil (5 to 80 mg), telmisartan (20 to 160 mg), eprosartan mesylate(75 to 600 mg), atenolol (25 to 200 mg), propranolol hydrochloride (10to 180 mg), metoprolol tartrate, succinate or fumarate (25 to 400 mg),nadolol (20 to 160 mg), betaxolol hydrochloride (10 to 40 mg),acebutolol hydrochloride (200 to 800 mg), pindolol (5 to 20 mg),bisoprolol fumarate (5 to 20 mg), nifedipine (15 to 100 mg), felodipine(2.5 to 20 mg), amlodipine besylate (2.5 to 20 mg), nicardipine (10 to40 mg), nisoldipine (10 to 80 mg), terazosin hydrochloride (1 to 20 mg),doxazosin mesylate (4 to 16 mg), prazosin hydrochloride (2.5 to 10 mg),or alfuzosin hydrochloride (10 to 20 mg).

In a further embodiment, the co-formulation can be formulated for oraladministration twice per 24 hours, for delayed release of the activeuntil gastric transit is complete, and for sustained release of activeover a period of 2 to 12 hours. Such drugs to treat hypertensioninclude, but are not limited to: enalapril maleate (1.25 to 20 mg),captopril (2 to 75 mg), lisinopril (5 to 20 mg), benzaeprilhydrochloride (5 to 40 mg), quinapril hydrochloride (5 to 40 mg),peridopril erbumine (2 to 4 mg), ramipril (1 to 10 mg), trandolapril (1to 4 mg), fosinopril sodium (5 to 40 mg), moexipril hydrochloride (2.5to 10 mg), losartan potassium (12.5 to 100 mg), irbesartan (37.5 to 300mg), valsartan (20 to 300 mg), candesartan cilexetil (2 to 32 mg),olmesartan medoxamil (2.5 to 40 mg), telmisartan (10 to 80 mg),eprosartan mesylate (37.5 to 300 mg), atenolol (12.5 to 100 mg),propranolol hydrochloride (5 to 90 mg), metoprolol tartrate, succinateor fumarate (12.5 to 200 mg), nadolol (10 to 80 mg), betaxololhydrochloride (5 to 20 mg), acebutolol hydrochloride (100 to 400 mg),pindolol (2.5 to 10 mg), bisoprolol fumarate (2.5 to 10 mg), nifedipine(7.5 to 50 mg), felodipine (1 to 10 mg), amlodipine besylate (1 to 10mg), nicardipine (5 to 20 mg), nisoldipine (5 to 40 mg), terazosinhydrochloride (1 to 10 mg), doxasoxin mesylate (2 to 8 mg), prazosinhydrochloride (1 to 5 mg), or alfuzosin hydrochloride (5 to 10 mg).

In another embodiment of the invention, K_(ATP) channel openers selectedfrom salts of compounds of Formulae I-VIII are co-formulated with adiuretic. Such co-formulations can be formulated for oral administrationonce per 24 hours, for delayed release of the active until gastrictransit is complete, and for sustained release of the active over aperiod of 2 to 24 hours. Such diuretics can include, but are not limitedto: amiloride hydrochloride (1 to 10 mg), spironolactone (10 to 100 mg),triamterene (25 to 200 mg), bumetanide (0.5 to 4 mg), furosemide (10 to160 mg), ethacrynic acid or ethacrynate sodium (10 to 50 mg), torsemide(5 to 100 mg), chlorthalidone (10 to 200 mg), indapamide (1 to 5 mg),hydrochlorothiazide (10 to 100 mg), chlorothiazide (50 to 500 mg),bendroflumethiazide (5 to 25 mg), hydroflumethiazide (10 to 50 mg),mythyclothiazide (1 to 5 mg), and polythiazide (1 to 10 mg).

In a further embodiment, the co-formulation can be formulated for oraladministration twice per 24 hours, for delayed release of the activeuntil gastric transit is complete, and for sustained release of theactive over a period of 2 to 12 hours. Such diuretics include, but arenot limited to: amiloride hydrochloride (0.5 to 5 mg), spironolactone (5to 50 mg), triamterene (12 to 100 mg), bumetanide (0.2 to 2 mg),furosemide (5 to 80 mg), ethacrynic acid or ethacrynate sodium (5 to 25mg), tosemide (2 to 50 mg), chlorthalidone (5 to 100 mg), indapamide(0.5 to 2.5 mg), hydrochlorothiazide (5 to 50 mg), chlorothiazide (25 to250 mg), bendroflumethiazide (2 to 12.5 mg), hydroflumethiazide (5 to 25mg), mythyclothiazide (0.5 to 2.5 mg), and polythiazide (0.5 to 5 mg).

In another embodiment of the invention, K_(ATP) channel openers selectedfrom salts of compounds of Formulae I-VIII are co-formulated with a drugto treat inflammation or pain. Such co-formulations can be formulatedfor oral administration once per 24 hours, for delayed release of theactive until gastric transit is complete, and for sustained release ofthe active over a period of 2 to 24 hours. Such drugs to treatinflammation or pain include, but are not limited to: aspirin (100 to1000 mg), tramadol hydrochloride (25 to 150 mg), gabapentin (100 to 800mg), acetaminophen (100 to 1000 mg), carbamazepine (100 to 400 mg),ibuprofen (100 to 1600 mg), ketoprofen (12 to 200 mg), fenprofen sodium(100 to 600 mg), flurbiprofen sodium or flurbiprofen (50 to 200 mg), orcombinations of these with a steroid or aspirin.

In a further embodiment, the co-formulation can be formulated for oraladministration twice per 24 hours, for delayed release of the activeuntil gastric transit is complete, and for sustained release of theactive over a period of 2 to 12 hours. Such drugs to treat inflammationor pain include, but are not limited to: aspirin (100 to 650 mg),tramadol hydrochloride (12 to 75 mg), gabapentin (50 to 400 mg),acetaminophen (50 to 500 mg), carbamazepine (50 to 200 mg), ibuprofen(50 to 800 mg), ketoprofen (6 to 100 mg), fenprofen sodium (50 to 300mg), flurbiprofen sodium or flurbiprofen (25 to 100 mg), or combinationsof these with a steroid or aspirin.

A method of inducing loss of greater than 25% of initial body fat in anoverweight or obese subject can be achieved by the prolongedadministration of an oral dosage form of a K_(ATP) channel openerselected from a salt of a compound of Formulae I-VIII.

A method of inducing loss of greater than 50% of initial body fat in anoverweight or obese subject can be achieved by the prolongedadministration of an oral dosage form of a K_(ATP) channel openerselected from a salt of a compound of Formulae I-VIII.

A method of inducing loss of greater than 75% of initial body fat in anoverweight or obese subject can be achieved by the prolongedadministration of an oral dosage form of a K_(ATP) channel openerselected from a salt of a compound of Formulae I-VIII.

A method of inducing preferential loss of visceral fat in an overweightor obese subject can be achieved by the prolonged administration of anoral dosage form of a K_(ATP) channel opener selected from a salt of acompound of Formulae I-VIII.

A method of inducing loss of body fat and reductions in circulatingtriglycerides in an overweight or obese subject can be achieved by theprolonged administration of an oral dosage form of a K_(ATP) channelopener selected from a salt of a compound of Formulae I-VIII.

In some embodiments, the invention provides a polymorph of a salt, whichsalt includes diazoxide and a cation selected from the group consistingof an alkali metal and a compound comprising a tertiary amine orquaternary ammonium group. In some embodiments, the cation is choline.

In some embodiments, the polymorph of diazoxide choline salt is of FormA having characteristic peaks in the XRPD pattern at values of two-theta(Cu Kα, 40 kV, 40 mA) at approximately 9.8, 10.5, 14.9, 17.8, 17.9,18.5, 19.5, 22.1, 22.6, 26.2, 29.6, and 31.2 degrees.

In some embodiments, the polymorph of diazoxide choline salt is of FormB having characteristic peaks in the XRPD pattern at values of two-theta(Cu Kα, 40 kV, 40 mA) at approximately 8.9, 10.3, 12.0, 18.3, 20.6,24.1, 24.5, 26.3, 27.1, and 28.9 degrees.

In some embodiments, the polymorph of diazoxide choline salt is of FormA having characteristic infrared absorbances at 2926, 2654, 1592, 1449,and 1248 cm⁻¹.

In some embodiments, the polymorph of diazoxide choline salt is of FormB having characteristic infrared absorbances at 3256, 2174, 2890, 1605,1463, and 1235 cm⁻¹.

In some embodiments, the polymorph of diazoxide includes potassium asthe cation.

In some embodiments, the polymorph of diazoxide potassium salt is ofForm A having characteristic peaks in the XRPD pattern at values oftwo-theta (Cu Kα, 40 kV, 40 mA) at approximately 6.0, 8.1, 16.3, 17.7,18.6, 19.1, 22.9, 23.3, 23.7, 24.7, 25.4, 26.1, 28.2, 29.6, and 30.2degrees.

In some embodiments, the polymorph of diazoxide potassium salt is ofForm B having characteristic peaks in the XRPD pattern at values oftwo-theta (Cu Kα, 40 kV, 40 mA) at approximately 8.5, 10.8, 16.9, 18.2,21.6, 25.5, 26.1, and 28.9 degrees.

In some embodiments, the polymorph of diazoxide potassium salt is ofForm C having characteristic peaks in the XRPD pattern at values oftwo-theta (Cu Kα, 40 kV, 40 mA) at approximately 5.7, 6.1, 17.9, 23.9,25.1, and 37.3 degrees.

In some embodiments, the polymorph of diazoxide potassium salt is ofForm D having characteristic peaks in the XRPD pattern at values oftwo-theta (Cu Kα, 40 kV, 40 mA) at approximately 5.7, 6.2, 8.1, 8.5,8.8, 16.9, 18.6, 23.2, 24.5, 25.8, and 26.1 degrees.

In some embodiments, the polymorph of diazoxide potassium salt is ofForm E having characteristic peaks in the XRPD pattern at values oftwo-theta (Cu Kα, 40 kV, 40 mA) at approximately 6.7, 7.1, 14.1, and21.2 degrees.

In some embodiments, the polymorph of diazoxide potassium salt is ofForm F having characteristic peaks in the XRPD pattern at values oftwo-theta (Cu Kα, 40 kV, 40 mA) at approximately 8.5, 9.0, 18.7, 20.6,23.5, 27.5, and 36.3 degrees.

In some embodiments, the polymorph of diazoxide potassium salt is ofForm G having characteristic peaks in the XRPD pattern at values oftwo-theta (Cu Kα, 40 kV, 40 mA) at approximately 5.2, 5.5, 13.1, 16.5,19.3, 22.8, 24.8, 26.4, 28.7, and 34.1 degrees.

In some embodiments, the polymorph of diazoxide potassium salt is ofForm A having characteristic infrared absorbances at 1503, 1374, 1339,1207, 1131, 1056, and 771 cm⁻¹.

In some embodiments, the polymorph of diazoxide potassium salt is ofForm B having characteristic infrared absorbances at 1509, 1464, 1378,and 1347 cm⁻¹.

In some embodiments, the polymorph of diazoxide potassium salt is ofForm C having characteristic infrared absorbances at 1706, 1208, 1146,and 746 cm⁻¹.

In some embodiments, the polymorph of diazoxide potassium salt is ofForm D having characteristic infrared absorbances at 1595, 1258, 1219,and 890 cm⁻¹.

In some embodiments, the polymorph of diazoxide potassium salt is ofForm E having characteristic infrared absorbances at 1550, 1508, 1268,1101, and 1006 cm⁻¹.

In some embodiments, the polymorph of diazoxide potassium salt is ofForm F having characteristic infrared absorbances at 1643, 1595, 1234,1145, and 810 cm⁻¹.

In some embodiments, the polymorph of diazoxide potassium salt is ofForm G having characteristic infrared absorbances at 1675, 1591, 1504,1458, 1432, 1266, 999, 958, 905, and 872 cm⁻¹.

In some embodiments, the polymorph of diazoxide choline salt is of FormA having characteristic peaks in the XRPD pattern substantially as shownin FIG. 16( a), and an NMR spectrum substantially as shown in FIG. 17(a).

In some embodiments, the polymorph of diazoxide choline salt is of FormB having characteristic peaks in the XRPD pattern substantially as shownin FIG. 16( c) and an NMR spectrum substantially as shown in FIG. 17(b).

In some embodiments, the polymorph of diazoxide potassium salt includesone or more of Forms A-G, wherein each of the Forms A-G hascharacteristic peaks in the XRPD pattern substantially as shown in FIG.18-19.

In some embodiments of the invention there are provided methods forproducing a diazoxide choline salt, which methods include suspendingdiazoxide in a solvent (e.g., alcohols such as methanol, i-BuOH, i-AmOH,t-BuOH, and the like, ketones, tetrahydrofuran, dimethylformamide,n-methylpyrrolidinone, and the like) and mixing with a choline salt(e.g., choline hydroxide), adding a co-solvent (e.g., MTBE, EtOA, IPA,c-Hexane, heptane, toluene, CH₂CL₂, dioxane, and the like) to thesuspension under conditions sufficient to cause formation andprecipitation of said diazoxide choline salt, and harvesting theprecipitate to provide the diazoxide choline salt.

In some embodiments, the solvent is tetrahydrofuran. In someembodiments, the solvent is 2-methyltetrahydrofuran (2-MeTHF).

In some embodiments, the diazoxide and the solvent are present at aratio of about 1 g diazoxide per 1 mL solvent to about 1 g diazoxide per5 mL solvent. In some embodiments, the diazoxide and the solvent arepresent at a ratio of about 1 g diazoxide per 3 mL solvent.

In some embodiments, the choline salt is a solution in MeOH. In someembodiments, the choline salt is choline hydroxide in about a 45%solution (e.g., 40%, 41%, 42%, 43%, 44%, 45%, 46%, 47%, 48%, 49%, 50%)in MeOH.

In some embodiments, the choline salt is added as 1 equivalent ofdiazoxide.

In some embodiments, the co-solvent is MTBE.

In some embodiments, the amount of co-solvent added is in a ratio to theamount of the solvent of about 3:14 (solvent:co-solvent) (e.g., 3:12,3:13, 3:14, 3:15, 3:16).

In some embodiments, the process of making polymorphs of diazoxidecholine salt includes the step of seeding with crystals of diazoxidecholine salt polymorph Form B prior to the harvesting step.

In some embodiments for the method for producing a diazoxide cholinesalt the salt includes polymorph Form B substantially free of polymorphForm A, the polymorph Form B having characteristic peaks in the XRPDpattern at values of two-theta (Cu Kα, 40 kV, 40 mA) at approximately8.9, 10.3, 12.0, 18.3, 20.6, 24.1, 24.5, 26.3, 27.1, and 28.9 degrees.

In some embodiments for the method of treating obesity orobesity-related morbidity in an obese subject, the compound is acompound of Formula V.

In some embodiments for the method of treating obesity orobesity-related morbidity in an obese subject, the compound is acompound of Formula VI.

In some embodiments for the method of treating obesity orobesity-related morbidity in an obese subject, the compound is acompound of Formula VII.

In some embodiments for the method of treating obesity orobesity-related morbidity in an obese subject, the compound is acompound of Formula VIII.

In some embodiments for the method of treating obesity orobesity-related morbidity in an obese subject, the method furthercomprises administering a drug selected from the group consisting ofSibutramine, Orlistat, Rimonabant, an appetite suppressant, anon-thiazide diuretic, a drug that lowers cholesterol, a drug thatraises HDL cholesterol, a drug that lowers LDL cholesterol, a drug thatlowers blood pressure, a drug that is an anti-depressant, a drug that isan anti-epileptic, a drug that is an anti-inflammatory, a drug that isan appetite suppressant, a drug that lowers circulating triglycerides,and a drug that is used to induce weight loss in an overweight or obeseindividual.

In some embodiments for the method of treating obesity orobesity-related morbidity in an obese subject, the method furthercomprises administering a pharmaceutically active agent other than theK_(ATP) channel opener. In some embodiments, the other pharmaceuticallyactive agent is an agent useful for the treatment of a conditionselected from the group consisting of obesity, prediabetes, diabetes,hypertension, depression, elevated cholesterol, fluid retention, obesityassociated co-morbidities, ischemic and reperfusion injury, epilepsy,cognitive impairment, schizophrenia, mania, and other psychoticcondition.

In some embodiments for the method for treating a subject suffering fromor at risk for Alzheimer's disease (AD), the method includesadministration to a subject a therapeutically effective amount of a saltof diazoxide including salts provided herein. In some embodiments forthe method for treating a subject suffering from or at risk for AD, themethod includes administration to a subject a therapeutically effectiveamount of a compound according to any of Formulae I-VIII. In someembodiments, the compound is diazoxide or a salt thereof.

AD is a neurodegenerative disorder neuropathologically characterized byabnormal accumulations of intracellular neurofibrilary tangles andextracellular amyloid plaques throughout cortical and limbic brainregions and the loss of synapses and neurons. AD is furthercharacterized by significant cognitive and memory impairment. β amyloidplaques form the β amyloid peptide, either 1-40 or 1-42 peptide, whichis released from amyloid precursor protein following cleavage by gammasecretase. In addition to forming plaques, the β amyloid peptides arecytotoxic either as the monomer or as a short-lived oligomericintermediate. β amyloid peptides (monomers, dimers or oligomers) can beidentified both in CSF (cerebrospinal fluid) and in serum. Amyloidangiopathy is characterized by Aβ deposition and may contribute to thecerebrovascular abnormalities that precede the onset of AD.

The invention will now be described with reference to the followingnon-limiting examples.

EXAMPLES A. Potassium ATP Channel Activator Containing Formulations

1. Compressed Tablet Formulations of Diazoxide Salt or Derivative

Diazoxide salt or a derivative thereof at about 15-30% by weight ismixed with hydroxypropyl methylcellulose at about 55-80% by weight,ethylcellulose at about 3-10 wt/vol % and magnesium stearate (aslubricant) and talc (as glidant) each at less than 3% by weight. Themixture is used to produce a compressed tablet as described in Reddy etal., AAPS Pharm Sci Tech 4(4):1-9 (2003). The tablet may be coated witha thin film as discussed below for microparticles.

A tablet containing 100 mg of diazoxide salt or a derivative thereofwill also contain approximately 400 mg of hydroxypropyl cellulose and 10mg of ethylcellulose. A tablet containing 50 mg of diazoxide salt or aderivative thereof will also contain approximately 200 mg ofhydroxypropyl cellulose and 5 mg of ethylcellulose. A tablet containing25 mg of diazoxide salt or a derivative thereof will also containapproximately 100 mg of hydroxypropyl cellulose and 2.5 mg ofethylcellulose.

2. Encapsulated Coated Microparticle Formulation of Diazoxide Salt orDerivative

Diazoxide salt or a derivative thereof is encapsulated intomicroparticles in accordance with well known methods (see, e.g. U.S.Pat. No. 6,022,562). Microparticles of between 100 and 500 microns indiameter containing diazoxide salt or a derivative thereof, alone or incombination with one or more suitable excipient, is formed with theassistance of a granulator and then sieved to separate microparticleshaving the appropriate size. Microparticles are coated with a thin filmby spray drying using commercial instrumentation (e.g. Uniglatt SprayCoating Machine). The thin film comprises ethylcellulose, celluloseacetate, polyvinylpyrrolidone and/or polyacrylamide. The coatingsolution for the thin film may include a plasticizer which may be castoroil, diethyl phthalate, triethyl citrate and salicylic acid. The coatingsolution may also include a lubricating agent which may be magnesiumstearate, sodium oleate, or polyoxyethylenated sorbitan laurate. Thecoating solution may further include an excipient such as talc,colloidal silica or of a mixture of the two added at 1.5 to 3% by weightto prevent caking of the film coated particles.

3. Formulations for Controlled Release of Diazoxide or Derivative

3.1. Formulation of a Tableted Form of Diazoxide or a Derivative forControlled Release

Prior to mixing, both the active ingredient and hydroxypropylmethylcellulose (Dow Methocel K4M P) are passed through an ASTM 80 meshsieve. A mixture is formed from 1 part diazoxide salt or a derivativethereof to 4 parts hydroxypropyl methylcellulose. After thorough mixing,a sufficient volume of an ethanolic solution of ethylcellulose as agranulating agent is added slowly. The quantity of ethylcellulose pertablet in the final formulation is about 1/10th part. The mass resultingfrom mixing the granulating agent is sieved through 22/44 mesh.Resulting granules are dried at 40° C. for 12 hours and thereafter keptin a desiccator for 12 hours at room temperature. Once dry the granulesretained on 44 mesh are mixed with 15% fines (granules that passedthrough 44 mesh). Talc and magnesium stearate are added as glidant andlubricant at 2% of weight each. A colorant is also added. The tabletsare compressed using a single punch tablet compression machine.

3.2. Formulation of a Compression Tableted Form of Diazoxide or aDerivative Thereof that Provides for Controlled Release.

Diazoxide salt or a derivative thereof at 20-40% weight is mixed with30% weight hydroxypropyl methylcellulose (Dow Methocel K100LV P) and20-40% weight impalpable lactose. The mixture is granulated with theaddition of water. The granulated mixture is wet milled and then dried12 hours at 110° C. The dried mixture is dry milled. Following milling,25% weight ethylcellulose resin is added (Dow Ethocel 10FP or Ethocel100FP) followed by 0.5% weight magnesium stearate. A colorant may alsobe added. The tablets are compressed using a single punch tabletcompression machine (Dasbach et al., Poster at AAPS Annual Meeting Nov.10-14 (2002)).

3.3. Formulation of a Compression Coated Tableted Form of Diazoxide or aDerivative Thereof that Provides for Controlled Release.

The core tablet is formulated by mixing either 100 mg of diazoxide saltor a derivative thereof with 10 mg of ethylcellulose (Dow Ethocel 10FP),or by mixing 75 mg of diazoxide or a derivative thereof with 25 mglactose and 10 mg of ethylcellulose (Dow Ethocel 10FP), or by mixing 50mg of diazoxide or a derivative thereof with 50 mg of lactose and 10 mgof ethylcellulose (Dow Ethocel 10FP). The core tablets are formed on anautomated press with concave tooling. The compression coating consistingof 400 mg of polyethylene oxide (Union Carbide POLYOX WSR Coagulant) isapplied and compressed to 3000 psi (Dasbach et al., Poster at AAPSAnnual Meeting Oct. 26-30 (2003)).

3.4. Formulation of a Controlled Release Tableted Form of DiazoxideCholine Salt

3.4.1 Controlled Release Formulations

Controlled release tableted formulations of diazoxide choline salt weredeveloped and investigated with respect to a variety of properties knownby those of skill in the pharmaceutical art relating to the manufactureof tableted formulations including, for example, ease and consistency ofmanufacture, appearance (e.g., sheen, compressibility, microscopicappearance), and dissolution properties (e.g., rate, order and extent ofdissolution). Tablets were produced individually on a press, where thefinal blend of diazoxide choline salt and excipient was weighed out tothe desired total tablet weight prior to compression. As shown in Table2, Formulations A-H, J, and L contained 50.0 mg diazoxide as the cholinesalt (i.e., 72.5 mg total diazoxide choline salt present), andFormulations I and K contained 200.0 mg diazoxide as the choline salt(i.e., 290.0 mg total diazoxide choline salt present). The manufactureof formulation L is exemplary of the manufacturing methods available tothe skilled artisan. For formulation L, diazoxide choline salt, talc,and approximately half of the colloidal silicon dioxide (Cab-o-sil) weremixed in a KG-5 mixer bowl with an impeller speed of about 300 rpm and achopper speed of about 3000 rpm for about 4 min. The mixture was passedthrough a co-mil equipped with a 024R screen, square-edged paddle, and0.175″ spacer. To this milled mixture in a 8-qt V-shell blender wasadded Emcompress through a #20 mesh hand screen with blending for about10 min. To this mixture was added PEO N750 and PEO 303, which had beenpassed through a #20 mesh hand screen, with blending for about 10 min.To this mixture was added Pruv and the remainder of the Cab-o-sil,having been passed through a #20 mesh hand screen, with blending forabout 5 min. The mixture was subjected to pressing (Manesty Beta Press)using 0.2220″×0.5720″ caplet shaped tooling (Set #21.)

TABLE 2 Exemplary Formulations for Diazoxide Choline salt. FormulationINGREDIENT Amount per tablet (mg) A. Diazoxide Choline 72.50 Kollidon SR100.0 PEO N303 25.00 Emcompress 50.50 Pruv 2.00 TOTAL WEIGHT 250.0 B.Diazoxide Choline 72.50 Methocel K100M 237.3 Emcompress 35.00 MagnesiumStearate 3.50 Cab-o-sil 1.75 TOTAL WEIGHT 350.0 C. Diazoxide Choline72.50 Kollidon SR 105.0 PEO N303 35.0 Emcompress 134.1 Pruv 3.50 TOTALWEIGHT 350.1 D. Diazoxide Choline 72.50 Methocel K100M 175.1 Emcompress97.30 Magnesium Stearate 3.50 Cab-o-sil 1.75 TOTAL WEIGHT 350.2 E.Diazoxide Choline 72.50 PEO N750 NF 105.0 PEO N303 NF 52.50 Emcompress116.6 Pruv 3.50 TOTAL WEIGHT 350.1 F. Diazoxide Choline 72.50 KollidonSR 105.0 PEO N303 7.00 Emcompress 162.1 Pruv 3.50 TOTAL WEIGHT 350.1 G.Diazoxide Choline 72.50 Methocel K100M 175.1 Emcompress 105.1 MagnesiumStearate 3.50 Cab-o-sil 1.75 TOTAL WEIGHT 358.0 H. Diazoxide Choline72.50 PEO N750 NF 105.0 PEO N303 NF 35.01 Emcompress 134.1 Pruv 3.50TOTAL WEIGHT 350.1 I. Diazoxide Choline 290.0 Methocel K100M 240.0Emcompress 258.0 Magnesium Stearate 8.00 Cab-o-sil 4.00 TOTAL WEIGHT800.0 J. Diazoxide Choline 72.50 PEO N750 NF 105.0 PEO N303 NF 52.50Emcompress 116.6 Pruv 3.50 Talc 3.50 Cab-o-sil 1.75 TOTAL WEIGHT 355.4K. Diazoxide Choline 290.0 PEO N750 NF 249.0 PEO N303 NF 124.5Emcompress 145.3 Pruv 8.30 Talc 8.30 Cab-o-sil 4.15 TOTAL WEIGHT 829.6L. Diazoxide Choline 72.51 PEO N750 NF 105.1 PEO N303 NF 52.54Emcompress 111.4 Pruv 3.50 Talc 3.50 Cab-o-sil 1.75 TOTAL WEIGHT 350.3

Microscopic observation of the tablets having Formulation A revealed agrainy texture of the diazoxide choline salt, and at the 29% loading ofdiazoxide choline salt in Formulation A the blend had poor flowcharacteristics. Accordingly, the loading of diazoxide choline wasreduced in Formulation B. A caplet shaped tooling, approximately 6 mm×15mm was found to result in acceptable tablet appearance, sheen, and easeof compressibility. However, Formulation B also exhibited poor blendflow.

Subsequent formulations (e.g., Formulations C-L) incorporated a millingstep of the diazoxide choline salt prior to incorporation into thetablet blend. A milling study was conducted using a test mill equippedwith different screen sizes to evaluate and determine a suitable millingprocess. Particle size was determined by visual comparison with 40 μmreference beads. As shown in Table 3, use of the 024R screen resulted inthe best recovery of API (i.e., “active pharmaceuticalingredient”=diazoxide as the diazoxide choline salt) providing thebroadest range of particle sizes. Accordingly, material milled through a024R screen was selected for subsequent formulation.

TABLE 3 Milling study for diazoxide choline salt prior to formulationAPI loaded/API recovered Particle size Screen/Paddle/Spacer/Speed (g)(μm) 018R/square/0.200″/80% 50.0/36.5 ~100-200 024R/square/0.200″/80%50.0/42.0 ~100-250 039R/square/0.200″/80% 50.0/42.0 ~150-250

3.4.2. Dissolution Studies

Dissolution of tablets with formulation as set forth in Table 4 wasinvestigated. One or more tablets (e.g., 1 or 2) of the indicatedtableted formulation were placed into a volume of buffer (e.g., 900 mL)with known buffer salt concentration (e.g., 0.05 M potassium phosphate,pH 8.6; 0.05 M potassium phosphate, pH 7.5), with or without surfactant(e.g., 0.05% CTAB), at a known temperature (e.g., 37° C.). Stirringconditions employed paddles at, for example, 50 RPM. Aliquots (e.g, 10mL) removed as a function of time were filtered (e.g., 0.45 μm GMFFilter) prior to analysis.

TABLE 4 Dissolution study for formulations of diazoxide choline salt(entries are % dissolved diazoxide) Time (hr) Entry Protocol 0 1 2 3 6 912 18 24 C a 0.0 2.9 4.5 5.7 9.4 13.0 16.6 D a 0.0 4.5 7.8 11.0 19.125.6 31.3 F b 29 36 44 54 G b 30 39 50 67 H b 50 75 91 101 I b 25 37 4857 J b 34 53 76 104 K b 25 43 64 94 24 41 62 92 Column 1: Entry fromTable 2. Column 2: Protocol a) 0.05M potassium phosphate, pH 8.6, 37°C.; Protocol b) 0.05M potassium phosphate, pH 7.5, 0.05% CTAB, 37° C.

The dissolution profile (i.e., % dissolved with time) of Proglycem®capsules (100 mg) and controlled-release tablet formulations ofdiazoxide as provided herein is shown in Table 5. The 50 mg tablet entryof Table 5 refers to Formulation J (Table 2) wherein talc and cab-o-silwas present at 2% and 1%, respectively. The 200 mg tablet entry of Table5 refers to Formulation K (Table 2) wherein talc and cab-o-sil arepresent at 2% and 1%, respectively

TABLE 5 Dissolution profile of 100 mg Proglycem ® (capsule) anddiazoxide choline salt (tablet) 200 mg tablet 50 mg tablet 100 mgcapsule diazoxide choline diazoxide choline Time (hr) Proglycem ® saltsalt 0 0 0 0 3 79 22 25 6 85 36 39 12 92 54 70 24 98 88 99

As evidenced by Table 5, the 100-mg Proglycem® capsule provides forfaster dissolution of diazoxide relative to tablets described herein.Approximately 79% diazoxide component of Proglycem® is recovered indissolution buffer after 3-hr. In contrast, the 50 and 200-mg tabletsdescribed in Table 5 dissolved at levels of 25% and 22%, respectively,at 3-hr. At 12-hr, 100-mg Proglycem® capsule dissolved at 92%, whereasthe 50 and 200-mg tablets dissolved at 70% and 54%, respectively.Approximately total dissolution is observed with 100-nmg Proglycem®capsule and 50-mg tablet at 24 hrs.

3.4.3. Excipient Compatibility Studies

Studies to determine the compatibility of diazoxide choline salt forvarious excipients are summarized in Table 6. Each mixture of excipient(100 mg) and diazoxide choline salt (100 mg) was made in acetonitrile to10 mL Samples were assayed immediately (i.e., “Initial” column of Table6) and at one-month storage under conditions a) 40° C./75% RH (relativehumidity), and b) 50° C.

TABLE 6 Excipient compatibility study for diazoxide choline salt. 1Month 1 Month Initial % 40° C./75% RH % 50° C. % Excipient (w/w) (w/w)(w/w) Hydroxyrpropylmethyl cellulose (HMPC) 98.8 101.4 96.6Hydroxypropylcellulose (HPC) 103.1 97.4 100.8 Ethylcellulose (EC) 90.6102.0 99.7 99.5* Methylcellulose (MC) 102.3 100.3 99.1 Carboxymethylcellulose Na (CMC Na) 96.8 101.6 99.5 Starch 1500 103.3 90.6 95.2Kollidon SR 81.2 99.8 100.9 100.2* Polyethyleneoxide N3 03 (PEO) 81.099.2 99.0 99.1* Dibasic Calcium Phosphate 98.4 102.9 102.2 SodiumStearyl Fumarate 98.9 101.1 103.7 Magnesium Stearate 100.1 101.4 99.3Colloidal Silicon Dioxide (Cab-o-sil) 99.0 99.4 104.5 MicrocrystallineCellulose 96.8 98.6 107.0 Lactose Monohydrate 94.5 99.2 103.0 Mannitol111.7 98.1 81.3 Diazoxide Control N/A 99.0 99.4 *Re-assayed

Initial low results for ethylcellulose, Kollidon SR, andpolyethyleneoxide were investigated by re-preparing samples of diazoxidecholine salt and excipient. As shown in Table 6, initial sample recovery(i.e., column “Recovery”) of the re-prepared samples indicates methodaccuracy.

No reportable impurities were detected in any sample, and no degradationwas observed in the samples stored for one-month with the exception ofmannitol wherein 81.3% of diazoxide was recovered after 1-month at 50°C.

3.4.3. Stability Studies for Diazoxide Choline Controlled ReleaseTablets

Studies to determine the stability of formulations of diazoxide cholinesalt as the controlled release tablet were conducted on sampleformulations as described in Table 2, results of which are shown inTable 7. In these studies, storage conditions were the following: a) 25°C./60% RH, and b) 40° C./75% RH. In Table 7, the term “Appearance”refers to the physical appearance of the tablets of the study. The term“Assay (%)” refers to the percentage of diazoxide choline salt assayedwith respect to nominal (i.e., 50 mg or 200 mg) content of the sampleThe term “Dissolution (%)” refers to the amount, expressed as apercentage, of diazoxide choline salt assayed by method describedherein.

TABLE 7 Stability Study for Formulations of Diazoxide Choline Salt. Timepoint Test Initial 1 month 2 months 50 mg tablet, Formulation J, 25°C./60% RH, Appearance White tablets White tablets White tablets Assay(%) 95.8 102.9 99.3 Dissolution (%):  3 hr 24 19 15  6 hr 41 30 27 12 hr62 48 52 24 hr 92 90 94 50 mg tablet, Formulation J, 40° C./75% RH,Appearance White tablets White tablets White tablets Assay (%) 95.8103.2 99.4 Dissolution (%):  3 hr 24 10 10  6 hr 41 22 21 12 hr 62 50 4624 hr 92 90 84 200 mg tablet, Formulation K, 25° C./60% RH AppearanceWhite tablets White tablets White tablets Assay (%) 95.9 100.9 100.3Dissolution (%):  3 hr 34 15 13  6 hr 53 31 30 12 hr 76 69 68 24 hr 104101 96 200 mg tablet, Formulation K, 40° C./75% RH Appearance Whitetablets White tablets White tablets Assay (%) 95.9 99.6 100.0Dissolution (%):  3 hr 34 10 10  6 hr 53 26 28 12 hr 76 60 61 24 hr 10490 90

3.5. A Controlled Release Dosage Form of Diazoxide or a DerivativeThereof Using an Osmotically Controlled Release System.

Diazoxide salt or a derivative thereof is formulated as an osmoticallyregulated release system. In general, two components, and an expandablehydrogel that drives release of the active drug is assembled withdiazoxide salt or a derivative thereof into a semipermeable bilaminateshell. Upon assembly a hole is drilled in the shell to facilitaterelease of active upon hydration of the hydrogel.

A dosage form adapted, designed and shaped as an osmotic delivery systemis manufactured as follows: first, a diazoxide salt or a derivativethereof composition is provided by blending together into a homogeneousblend of polyethylene oxide, diazoxide salt or a derivative thereof andhydroxypropyl methylcellulose. Then, a volume of denatured anhydrousethanol weighing 70% of the dry mass is added slowly with continuousmixing over 5 minutes. The freshly prepared wet granulation is screenedthrough a 20 mesh screen through a 20 mesh screen, dried at roomtemperature for 16 hours, and again screened through a 20 mesh screen.Finally, the screened granulation is mixed with 0.5% weight of magnesiumstearate for 5 minutes.

A hydrogel composition is prepared as follows: first, 69% weight ofpolyethylene oxide weight, 25% weight of sodium chloride and 1% weightferric oxide are separately screened through a 40 mesh screen. Then, allthe screened ingredients are mixed with 5% weight of hydroxypropylmethylcellulose to produce a homogeneous blend. Next, a volume ofdenatured anhydrous alcohol equal to 50% of the dry mass is added slowlyto the blend with continuous mixing for 5 minutes. The freshly preparedwet granulation is passed through a 20 mesh screen, allowed to dry atroom temperature for 16 hours, and again passed through a 20 meshscreen. The screened granulation is mixed with 0.5% weight of magnesiumstearate for 5 minutes (see U.S. Pat. No. 6,361,795 by Kuczynski, etal.).

The diazoxide salt composition, or a derivative thereof, and thehydrogel composition, are compressed into bilaminate tablets. First thediazoxide salt or a derivative thereof composition is added and tamped.The hydrogel composition is then added and the laminae are pressed undera pressure head of 2 tons into a contacting laminated arrangement.

The bilaminate arrangements are coated with a semipermeable wall (i.e.thin film). The wall forming composition comprises 93% cellulose acetatehaving a 39.8% acetyl content, and 7% polyethylene glycol. The wallforming composition is sprayed onto and around the bilaminate.

Finally, an exit passageway can be drilled through the semipermeablewall to connect the diazoxide salt or a derivative thereof drug laminawith the exterior of the dosage system. Residual solvent is removed bydrying at 50° C. and 50% humidity. The osmotic systems are dried at 50°C. to remove excess moisture (see U.S. Pat. No. 6,361,795 by Kuczynski,et al.).

4. Preparation of Diazoxide Salts

4.1. Preparation of the Sodium Salt

The sodium salt of diazoxide was prepared by dissolving 300 mg ofdiazoxide in approximately 45 mL methyl ethyl ketone (MEK). Thediazoxide/MEK solution was heated at 75° C. on an orbital shaker toensure dissolution. To the solution was added 1.3 mL of 1M NaOH (1 molarequivalent). The combined solutions were heated at 75° C. forapproximately 30 minutes and allowed to cool to room temperature. Themixture was concentrated under reduced pressure, and dried in vacuo at55° C. and 30 in. Hg. Elemental analysis: Calculated, 38.03% C, 2.39% H,11.09% N and 9.1% Na; Found, 38.40% C, 2.25% H, 10.83% N and 7.4% Na.

A sodium salt of diazoxide was also prepared by dissolving 300 mg ofdiazoxide in approximately 45 mL acetonitrile. Thediazoxide/acetonitrile solution was heated at 75° C. on an orbitalshaker to ensure dissolution. To the solution was added 1.3 mL of 1MNaOH (approximately 1 molar equivalent). The combined solutions wereheated at 75° C. for approximately 30 minutes and allowed to cool toroom temperature. The mixture was concentrated under reduced pressure,and dried in vacuo at 55° C. and 30 in. Hg.

4.2. Preparation of the Potassium Salt

The potassium salt of diazoxide was prepared by dissolving 300 mg ofdiazoxide in approximately 45 mL methyl ethyl ketone (MEK). Thediazoxide/MEK solution was heated at 75° C. on an orbital shaker toensure dissolution. To the solution was added approximately 1.3 mL of 1M KOH (1 molar equivalent) and the solution was returned to the orbitalshaker, heated at 75° C. for approximately 30 minutes and allowed tocool to room temperature. The solvent was removed under reduced pressureand the solid was dried in vacuo at 55° C. and 30 in. Hg. Elementalanalysis: Calculated, 33.59% C, 2.81% H, 9.77% N and 13.63% K; Found,34.71% C, 2.62% H, 9.60% N and 10.60% K.

The potassium salt of diazoxide was also prepared by dissolving 300 mgof diazoxide in approximately 45 mL tetrahydrofuran (THF). Thediazoxide/THF solution was heated at 75° C. on an orbital shaker toensure dissolution. To the solution was added approximately 1.3 mL of 1M KOH (1 molar equivalent) and the resulting solution was returned tothe orbital shaker, heated at 75° C. for approximately 30 minutes andallowed to cool to room temperature. THF was removed under reducedpressure and the solid was dried in vacuo at 55° C. and 30 in. Hg.

4.3. Preparation of Choline Salt

4.3.1. Preparation of the Choline Salt: Proof of Concept

The choline salt of diazoxide was prepared by dissolving 300 mg ofdiazoxide in approximately 45 mL methyl ethyl ketone (MEK). Thediazoxide/MEK solution was heated at 75° C. on an orbital shaker toensure dissolution. To the solution was added approximately 315 mg of 50wt. % of choline hydroxide (1 molar equivalent) and the solution wasreturned to the orbital shaker and stirred at 75° C. for approximately30 minutes. The solvent was removed under reduced pressure, and thesolid was dried in vacuo at 55° C. and 30 in. Hg. Elemental analysis:Calculated, 46.77% C, 5.86% H, and 12.00% N; Found, 46.25% C, 6.04% H,and 12.59% N.

4.3.2. Process of Preparation of the Choline Salt

An investigation of the minimum solvent volumes required to optimize theformation of the diazoxide choline salt was performed in MeCN, THF, MEK,and 2-MeTHF on small scale in the presence and absence of MTBE. Analysesby ¹H NMR and XRPD of all reactions were consistent with the assignedstructure and Form B of diazoxide choline salt.

4.3.2.1. Solvent Efficiency Study with Single Solvent System.

Investigation of single solvent system synthesis of diazoxide cholinesalt using solvents THF, MEK, and 2-MeTHF were conducted, results ofwhich are shown in Table 8. These results suggest that precipitation canbe achieved with a single solvent system in THF or 2-MeTHF. The bestresults, (i.e., Table 8 entries 14-17) obtained with 2-MeTHF. However,it was observed that in 2-MeTHF at 3-volumes and above, completedissolution was not achieved after addition of choline hydroxide.

TABLE 8 Solvent Efficiency of a Single Solvent System Entry SolventSolvent volume Yield (%) Polymorphic Form 1 THF 1.0  7 B 2 THF 2.0 n/an/a 3 THF 3.0 20 B 4 THF 4.0 28 B 5 THF 5.0 32 B 6 THF 6.0 42 B 7 THF7.0 53 B 8 THF 8.0 50 B 9 MEK 4.0 n/a n/a 10 2-MeTHF 1.0 n/a n/a 112-MeTHF 2.0 48 B 12 2-MeTHF 3.0 41 B 13 2-MeTHF 4.0 48 B 14 2-MeTHF 5.071 B 15 2-MeTHF 6.0 71 B 16 2-MeTHF 7.0 72 B 17 2-MeTHF 8.0 67 B n/a:Samples were either seen to form a gum, or no precipitate was observed.

4.3.2.2. Solvent Efficiency Study with Binary Solvent Systems.

The addition of a second solvent at 1-20 volumes (i.e., 1-8, 9, 10, 11,12, 13, 14, 15, 16, 17, 18, 19, 20) to the solvent volumes of Table 8was envisaged to optimize diazoxide salt production yields. Results ofinvestigation of methods to enhance precipitation and increase yieldemploying binary solvent system synthesis of diazoxide choline saltusing solvents THF, MeCN, MEK, and 2-MeTHF and co-solvent MTBE are shownin Table 9. Optimum conditions for the preparation of the diazoxidecholine salt obtained with 1-3 volumes of THF (see Table 8, entries1-3), with 3 volumes being the ratio of choice to eliminate excessivedrag during stirring of slurry in large scale production.

TABLE 9 Solvent Efficiency of a Binary Solvent System Solvent Co-SolventPolymorphic Entry Solvent Volume (12 Vol) Yield (%) Form 1 THF 1.0 MTBE96 B 2 THF 2.0 MTBE 96 B 3 THF 3.0 MTBE 91 B 4 THF 4.0 MTBE 90 B 5 THF5.0 MTBE 84 B 6 THF 6.0 MTBE 90 B 7 THF 7.0 MTBE 94 B 8 THF 8.0 MTBE 94B 9 MeCN 1.0 MTBE 92 B 10 MeCN 2.0 MTBE 90 B 11 MeCN 3.0 MTBE 87 B 12MeCN 4.0 MTBE 79 B 13 MeCN 5.0 MTBE 79 B 14 MeCN 6.0 MTBE 75 B 15 MeCN7.0 MTBE 70 B 16 MeCN 8.0 MTBE 74 B 17 MEK 4.0 MTBE 83 B 18 2-MeTHF 1.0MTBE 97 B 19 2-MeTHF 2.0 MTBE 94 B 20 2-MeTHF 3.0 MTBE 94 B 21 2-MeTHF4.0 MTBE 95 B 22 2-MeTHF 5.0 MTBE 97 B 23 2-MeTHF 6.0 MTBE 94 B 242-MeTHF 7.0 MTBE 97 B 25 2-MeTHF 8.0 MTBE >99 B

4.3.2.3. Co-Solvent Efficiency and Optimization with MTBE.

Results of optimization of co-solvent (MTBE) volume in the presence of 3volumes of solvent are shown in Table 10.

TABLE 10 Co-Solvent Efficiency and Optimization with MTBE Entry SolventMTBE Volume Yield (%) Form 1 MeCN 8.0 76 B 2 MeCN 14.0 85 B 3 THF 6.0 93B 4 THF 8.0 93 B 5 THF 14.0 98 B

Optimized conditions for the production of the choline salt of diazoxidein a binary solvent system obtained with 3:14 (i.e., 1:4.7) THF/MTBE(v/v).

4.3.2.4. Optimization of Cooling Profile.

Optimization for the controlled precipitation of diazoxide choline saltwas conducted by varying the cooling temperatures and hold times.Reactions were carried out in MeCN, THF, and 2-MeTHF (3 vol) with MTBE(14 vol). The results are presented in Table 11. These results suggestthat optimum crystal growth was achieved with cooling to 0-5° C. foreight hours in THF and 2-MeTHF (Entries 5-6) when compared to those inMeCN (Entries 1, 2, and 4). These results however did not indicateenhanced precipitation when compared to previous studies carried out inTHF and 2-MeTHF when the slurries were allowed to stir for two hours. Inaddition no notable improvements were observed for the additionalcooling to −15° C. or with the filtration at ambient temperature.

TABLE 11 Cooling Profile Optimization for Controlled PrecipitationCooling Temp. Hold Time Entry Solvent (° C.) (hr) Yield (%) Form 1 MeCN0-5 2 89 B 2 MeCN −10 → −15 2 85 B 3 THF RT 2 80 B 4 MeCN RT 2 85 B 5THF 0-5 8 94 B 6 2-MeTHF 0-5 8 95 B

4.3.2.5. Optimization of Rate of Co-Solvent Addition.

The effect of rate and hold time of co-solvent addition on theprecipitation of diazoxide choline salt was examined. The reactions werecarried out on 500 mg scale in three volumes of primary solvent and 14volumes of MTBE (THF and 2-MeTHF) or 1:3 volume ratio (MeCN). Additionof the co-solvent was made dropwise in one portion for Entry 1, and forEntries 2 and 3 the co-solvent was added at a rate of 1 mL/20 minutesfor a total hold time of 140 minutes for the addition of 7 mL. Theresults, presented in Table 12, indicate that optimum precipitation ofdiazoxide choline salt was achieved by the addition of MTBE with 20minute hold times in between additions in THF or 2-MeTHF (entries 2-3).

TABLE 12 Optimization of Rate of Co-solvent addition Hold Time AdditionRate Entry Solvent (min) (mL/min) Yield (%) Form 1 MeCN 60 n/a 87 B 2THF 140 1.0/20 96 B 3 2-MeTHF 140 1.0/20 96 B

4.3.2.6. Thermal Stability Study.

The stability of the isolated diazoxide choline salt was investigated todetermine optimal drying conditions to minimize residual solventswithout degradation. Samples of the diazoxide choline salt prepared inTHF and MTBE (Entries 1-4) were dried under vacuum at 40° C. for variousdurations on small scale (100 mg). The study was then carried out on 5 gscale (Entry 5) at 30° C. for eight hours under vacuum to showreproducibility on large scale. The results are presented in Table 13.Analyses of the samples of Table 13 by ¹H NMR, XRPD, and HPLC indicatedthat elevated temperatures and prolonged drying times did not degradethe compound or affect the form of the compound.

TABLE 13 Thermal Stability Study OVI Drying Temp (THF/MTBE) PolymorphicEntry (C.) Time (h) (ppm) Form 1 RT 12 376/248 B 2 40 20 317/160 B 3 4028 276/127 B 4 40 42 268/128 B 5 30 8 241/509 B

4.3.2.6. Demonstration of 50-g Scale synthesis of Diazoxide Choline Saltin THF.

The preparation of the diazoxide choline salt was carried out in abinary-solvent system of THF and MTBE with a ratio of 1:4.7(solvent/co-solvent) volumes and cooling to 0-5° C. for two hours withstirring. A demonstration run for the large scale production utilizingthe modified procedure was carried out on 50 g-scale. Diazoxide (50 g)as a hot (62° C.) suspension in THF (140 mL) was treated with cholinehydroxide (45% solution in MeOH, 1.0 equiv) added (2 mL/min) over 30minutes. The resulting solution was stirred for 30 minutes, followed bycooling to 52° C. for the addition of MTBE (14 vol) over 45 minutes. Onaddition of two volumes of co-solvent precipitation was observed. Theresultant slurry was then allowed to cool naturally to ambienttemperature followed by further cooling to 0-5° C. with an ice/waterbath and an additional 2 hr stirring. The precipitate was isolated byvacuum filtration, and the filter cake rinsed with ice cold MTBE (˜50mL) and dried under vacuum at room temperature for 12 hours. OVIanalysis indicated that THF levels were above the recommended ICHguidelines (799 ppm) and the material was returned to the oven at 30° C.for eight hours to give diazoxide choline salt [70.87 g, 97% yield, 97%purity AUC] as a white crystalline solid. Analyses by ¹H NMR, XRPD, andHPLC were consistent with the assigned structure of Form B whilemaintaining a high purity with good yield.

4.3.2.7. Demonstration of 50-g Scale Synthesis of Diazoxide Choline Saltin 2-MeTHF.

A 50-g level synthesis was duplicated in 2-MeTHF/MTBE as an alternativeto THF/MTBE for the large scale production of the diazoxide choline saltdescribed above. No issues or concerns arose during the reaction otherthan complete dissolution was not achieved after addition of the cholinehydroxide. A white precipitate formed after addition of the co-solvent,which was isolated via vacuum filtration and dried under vacuum at 30°C. for eight hours to give diazoxide choline salt [71.51 g, 97% yield]as a white crystalline solid. Analyses by 1H, XRPD, and HPLC wereconsistent with the assigned structure and Form B. OVI analysis showed2-MeTHF and MTBE levels of 125 and 191 ppm, respectively, which werebelow the ICH guidelines.

4.3.2.8. 250-g Scale Synthesis of Diazoxide Choline Salt in THF.

Larger scale preparation of the diazoxide choline salt was carried outin a binary-solvent system of THF and MTBE with a ratio of 1:4.7 (v/v)volumes. Diazoxide as a hot (62° C.) suspension in THF (745 mL) wastreated with choline hydroxide as a 45% solution in MeOH (1.0 equiv)added over 30 minutes. The resulting solution was stirred for 30minutes, followed by cooling to 52° C. for the addition of MTBE (14 vol)over 45 minutes. On addition of two volumes of co-solvent, precipitationwas observed. The resultant slurry was then allowed to cool naturally toambient temperature followed by cooling to 0-5° C. with an ice/waterbath. The precipitate was isolated by vacuum filtration, and the filtercake rinsed with ice cold MTBE (approximately 250 mL) and dried undervacuum at 30° C. for 38 hours to give diazoxide choline salt (350.28 g,97.7% yield) as a white crystalline solid. Analyses by ¹H NMR, XRPD, andHPLC were consistent with the assigned structure of Form B whilemaintaining a high purity with good yield.

4.3.2.9. 2-kg Scale Synthesis of Diazoxide Choline Salt in THF.

A 12-L reaction flash was charged with 2.0 kg diazoxide and 5.0-L THFwith stirring and heating to 55° C. Choline hydroxide (45% solution inmethanol, 2.32 L) was added dropwise to this reaction mixture over about2.5 hr with stirring. The temperature was maintained at 60±5° C. Afteraddition of choline hydroxide, stirring was continued for about 30 min.The reaction mixture was clarified by in-line 10 micron filtration upontransfer to a 22-L reaction flask pre-charged with 2-L pre-filtered THF,into which was added 10-L pre-filtered MTBE dropwise. This reactionmixture was transferred to another flask which was then charged with anadditional 30-L pre-filtered MTBE dropwise, with adjustment oftemperature to <5° C. and stirring for about 2 hr. Diazoxide cholinesalt was recovered by vacuum filtration to afford 2.724 kg (94%)diazoxide choline salt (99.8%, HPLC purity), confirmed by ¹H NMR, IR,and UV/Visible analysis.

4.4. Preparation of the Hexamethyl Hexamethylene Diammonium HydroxideSalt

The hexamethyl hexamethylene ammonium salt of diazoxide was prepared bydissolving 50 mg of diazoxide in approximately 7.5 mL methyl ethylketone (MEK). The diazoxide/MEK solution was heated at 75° C. on anorbital shaker to ensure dissolution. To the solution was addedapproximately 2.17 mL of 0.1M hexamethyl hexamethylene ammoniumhydroxide solution (1 molar equivalent) and the solution was stirred at75° C. for an additional 10 minutes and then cooled to room temperatureat the rate of 30° C./h. The solvent was removed under reduced pressure,and the solid was dried in vacuo at 55° C. and 30 in. Hg.

4.5. Failure to Obtain Salts of Diazoxide and Derivatives

4.5.1. Failure to Obtain Salts from Alkali Metal Hydroxides

U.S. Pat. No. 2,986,573 (“the '573 patent”) describes the synthesis ofdiazoxide metal salts in aqueous or non-aqueous solutions in thepresence of an alkali metal alkoxide. According to the '573 patent,diazoxide can be dissolved in an alkali metal solution, and the saltobtained upon evaporation. Also described is a method for forming saltsfrom non-aqueous media wherein diazoxide and sodium methoxide aredissolved in anhydrous methanol and the solvent is evaporated to obtainthe sodium salt of diazoxide as a white solid.

Attempts were made to prepare a diazoxide salt from alkali metalhydroxides by the method described in the '573 patent. Salt preparationwas carried out in aqueous media by dissolving diazoxide in a basicsolution 1M NaOH, followed by evaporation of the solvent. A solid wasobtained and analyzed by XRPD (X-Ray Powder Diffraction) and NMR.However, this analysis confirmed that the solid obtained was thediazoxide starting material and not a salt.

Salt preparation was carried out in non-aqueous media by dissolvingdiazoxide in anhydrous methanol in the presence of either sodiummethoxide or potassium methoxide and stirring the mixture at 60° C. for15 minutes. The mixture was then cooled to room temperature whilestirred. After approximately two hours, a solid was recovered, isolatedby filtration, and dried in vacuo. Analysis by XRPD confirmed that thesolid obtained was the diazoxide starting material and not a salt.

4.5.2. Preparation of Salts in Methanol or Ethanol According to the '573patent

The preparation of diazoxide salts in methanol and ethanol was attemptedusing 22 different counter-ions according to the methods described bythe '573 patent. For example, 20 mg of diazoxide was dissolved in 5 mLof ethanol and stirred and heated to ensure dissolution of thediazoxide. To the stirred solution was added approximately 1 molarequivalents of sodium methoxide. The solution was stirred forapproximately 10-15 minutes at 60° C., and cooled to room temperaturefor approximately 2 hours. The resulting solid precipitate wasconcentrated under a nitrogen stream, and collected by filtration. Theproduct was dried in vacuo and analyzed by XRPD. As shown in FIG. 15,XRPD of the solids collected from the sodium methoxide experiment, (aswell as the solids collected from the potassium methoxide experiment,which was run under similar conditions), revealed that the solid was thediazoxide starting material and that no sodium or potassium salt wasprepared. In FIG. 15D is the XRPD pattern of free form diazoxide; inFIG. 15B is the XRPD pattern of the product of potassium methoxide inmethanol, and in FIG. 15C is the XRPD pattern of the product of sodiummethoxide in methanol.

Although not wishing to be bound by any theory, it is believed that apossible explanation for the failure to prepare diazoxide salts by themethods of the '573 patent (i.e., in the presence of alcohols, e.g.,methanol or ethanol), is that the alcohol may have an effect on thestability of the alkali salts of diazoxide. This was supported by UVspectroscopy analysis of alkali salts of diazoxide (sodium andpotassium). Both the sodium and potassium salts of diazoxide weresynthesized by reacting diazoxide with NaOH or KOH in MEK. Elementalanalysis, NMR and XRPD confirmed that the salts were made.

Upon dissolution of the sodium or the potassium salt in acetonitrile,the UV spectrum shows a shift into the red region of the spectrum forthe λ_(max) from approximately 268 nm for the free form of diazoxide toapproximately 298 nm for both the potassium and sodium salt (see FIG.1). Similarly, these salts can be stabilized in aqueous solutions byelevating the pH to above 9.0. A similar shift in the absorption maximumat pH 9.0 is measured using UV spectroscopy. Subsequent adjustment ofthe pH from greater than 9.0 to less than 6.2 results in the hydrolysisof the salt as measured by the recovery of the UV absorption pattern ofthe diazoxide free base (see FIG. 2). In contrast, when the sodium orthe potassium salt is dissolved in methanol, the UV-Vis spectrum of thesalt was identical to that of the diazoxide starting material, (see FIG.3).

These results demonstrate that using methanol as the solvent isincompatible with the synthesis of alkali salts of diazoxide. Inaddition, the results show that isolation of an alkali metal salt ofdiazoxide (or any other salt) in the presence of an alcohol, such asmethanol, may not be possible.

4.5.3. Failure to Obtain Salts from Acidic Counter Ions

Salt formation with diazoxide was also attempted using acidiccounter-ions, such as, for example, hydrochloric acid, maleic acid,sulfuric acid, phosphoric acid, sulfamic acid, acetic acid, citric acid,lactic acid, tartaric acid, malonic acid, methanesulfonic acid,ethanesulfonic acid, benzenesulfonic acid, p-toluenesulfonic acid,cyclohexylsulfamic acid, fumaric acid, benzoic acid, undecylenic acid,salicylic acid and quinic acid. However, no salt formation was observedfor any acid in any solvent.

For example, 100 mg of diazoxide was dissolved in 50 mL of acetone andheated to 35° C. To a stirred solution was added approximately 4.65molar equivalents of HCl. The reaction mixture was allowed to cool toroom temperature for approximately 3 hours, with no precipitationobserved. Solvent was removed in vacuo. The resulting solid was analyzedby XRPD, and the observed XRPD pattern was consistent with the free formdiazoxide starting material. In all cases, the attempted synthesis ofdiazoxide salts from acids was unsuccessful in all solvents attempted.

4.6. Preparation of Salts of Compounds of Formulae V-VIII

The chloride salt of 3-amino-4-methyl-1,2,4-benzothiadiazine-1,1-dioxideis prepared dissolving approximately 300 mg (1.4 mmol) in 45 mLacetonitrile. The mixture is heated to approximately 75° C. and stirredfor 30 min. To the stirred solution, approximately 1 molar equivalent ofHCl is added dropwise, and stirred for approximately 30 min at 75° C.The mixture is cooled to room temperature and the solvent is removedunder reduced pressure, affording the chloride salt as a solid.

The sodium salt of 3-amino-4-methyl-1,2,4-benzothiadiazine-1,1-dioxideis prepared dissolving approximately 300 mg (1.4 mmol) in 45 mLacetonitrile. The mixture is heated to approximately 75° C. and stirredfor 30 min. To the stirred solution, approximately 1 molar equivalent ofNaOH is added dropwise, and stirred for approximately 30 min at 75° C.The mixture is cooled to room temperature and the solvent is removedunder reduced pressure, affording the sodium salt as a solid.

5. Characterization of Prepared Diazoxide Salts

Synthesis of the desired salts was confirmed by X-Ray Powder Diffraction(XRPD), UV-Vis spectroscopy, and NMR. All spectra were compared with thespectra of the free form diazoxide (i.e., not a salt). Differentialscanning calorimetry (DSC), thermal gravimetric analysis (TGA), FTIR(Fourier transform infrared spectroscopy), NMR, UV-vis spectroscopy andmoisture sorption analysis were also performed.

5.1. Experimental Procedures

DSC analysis were conducted with a Mettler 822 DSC, by measuring theamount of energy released by a sample, as the sample was heated from 30°C. to between 300-500° C. at a rate of 10° C./min. Typical applicationsof DSC analysis include determination of melting point temperature andthe heat of melting; measurement of the glass transition temperature;curing and crystallization studies; and identification of phasetransformations.

TGA measurements were conducted with a Mettler 851 SDTA/TGA, bymeasuring weight loss as a function of increasing temperature, as thesamples were heated from 30° C. to 230° C. at a rate of 10° C./min. TheTGA can be used to analyze desorption and decomposition behavior,characterize oxidation behavior, set burnout or conditioning parameters(temperature/ramp rate/time), and determine chemical composition.

XRPD samples were analyzed with a Shimadzu XRD-6000 system, using a CuKa, 40 kV, 40 mA X-ray tube. The divergence and scatter slits were 1.00deg, and the receiving slit was 0.30 mm. Samples were continuouslyscanned at a range of 3.0-45.0 deg, with a step size of 0.04 deg., at ascan rate of 2 deg/min.

Fourier Transform Infrared Spectroscopy was measured with aThermo-Nicolet Avatar 370 with a Smart Endurance Attenuated TotalReflection (ATR) attachment. Compressed samples were analyzed, withcorrections for background noise being made. Using the IR spectrum,chemical bonds and the molecular structure of organic compounds can beidentified. Attenuated total reflectance (ATR) allows for the analysisof thin films, organic and inorganic, in areas as small as 10-15microns.

Nuclear Magnetic Resonance (NMR) was performed with a 400 MHz BrukerAvance with a 4 mm CP/MAS H-X probe. Acquisition of ¹H NMR spectra wereperformed by taking between 5-10 mg of the sample, dissolved inapproximately 0.78 mL of DMSO-d₆. Spectra were acquired with either 16or 32 scans, using a pulse delay of 1.0 sec, with a 10 μsec (30°) pulsewidth.

UV spectroscopy was performed with a Perkin-Elmer Lambda 25spectrometer. Samples were dissolved in acetonitrile, water and a buffersystem having a pH between 5.6 and 10. Spectra were acquired between 340and 190 nm, using a 1 cm path length with background correction.

Moisture Sorption Analysis was performed with a Hiden IGAsorp MoistureSorption Instrument. Samples were first dried at 0% relative humidity at25° C. until an equilibrium weight was reached, or for a maximum of 4hours. Samples were then subjected to an isothermal (25° C.) scan from10-90% relative humidity in steps of 10%. The samples were allowed toequilibrate to an asymptotic weight at each point for a maximum of 4hours. Following absorption, a desorption scan from 85% relativehumidity (at 25° C.) was run in steps of −10%, again allowing a maximumof 4 hours for the samples to equilibrate. The resulting samples afterdesorption were dried at 80° C. for two hours and analyzed by XRPD.

5.2. Free Form Diazoxide Characterization

The free form of diazoxide was characterized by XRPD, differentialscanning calorimetry (DSC), thermal gravimetric analysis (TGA), moisturesorption, ¹H NMR, FTIR and UV-vis spectroscopy to provide a baseline forcomparison with the salts. Free form diazoxide is highly crystalline, asshown by the XRPD pattern. (See FIG. 4( a)). The DSC shows a largeendothermic event at 330° C., and TGA shows that the free form ofdiazoxide is anhydrous, where diazoxide shows no weight loss below 200°C., and a weight loss of only 0.2% below 230° C. Moisture absorption ofthe free form diazoxide shows the material to be non-hygroscopic.Absorption of water by diazoxide was tested at between 0-90% relativehumidity (RH) at 25° C., showing absorption of approximately 0.04 wt %at 60% RH and 0.20 wt % at 90% RH. The molecule does not form a stablehydrate, as shown by the lack of hysteresis during desorption.Additionally, the XRPD pattern for the diazoxide before and afterabsorption of water indicate the same crystalline form.

UV-vis spectroscopy measurements taken of the free form diazoxide inneutral aqueous solution show a λmax at approximately 268 nm. Inacetonitrile, the λmax was 264 nm, demonstrating a small solvatochromicshift. As shown in FIG. 2, as pH increased the λmax also increased, fromapproximately 265 nm to approximately 280 nm, due to a change in theelectronics of the molecule.

Studies were also conducted to evaluate the likelihood for conversionand degradation under thermal stress. Samples were heated in a closedenvironment, protected from light, at 60° C. for approximately 14 days.The diazoxide showed no conversion or degradation at 7 days or 14 days.Diazoxide samples were found to be consistent with the starting materialwith respect to XRPD and DSC.

Slurry studies to determine propensity of inter-conversion of the solidform were conducted on the free form diazoxide at room temperature, inthe absence of light, using water, isopropyl alcohol, dichloromethaneand toluene. Approximately 20 mg of the free form diazoxide was stirredfor 14 days. Analysis by XRPD, DSC and HPLC were consistent with thestarting material, indicating that the free form of diazoxide did notconvert to alternate crystal forms.

5.3. Characterization of Sodium Diazoxide Salt

The XRPD pattern of the sodium salt of diazoxide was analyzed, showingthe material to be crystalline. (See FIG. 4( d)). The DSC analysisrevealed a major exothermic event at 448° C. Small transitions below400° C. are likely due to sample imperfections. TGA analysis showedweight loss of 0.2% and 0.03% below 120° C., which may be the result ofbound solvent. Moisture absorption performed from 0-90% relativehumidity at 25° C. showed the material to be hygroscopic as the sampledeliquesced at 90% relative humidity. The sample absorbed 1.2 wt % ofwater at 60% RH, and 6.6 wt % water at 80% RH. Hysteresis was observedupon desorption at 65 and 55% RH, indicating possible hydrate formation.(Possible hydrate formation was noted, although the amount of waterabsorbed was less than 0.5 mole). ¹H NMR showed a chemical shift in thearomatic and methyl resonances of the sodium salt, as expected due tochanges to the aromatic system. See FIG. 5 showing NMR spectrum for thefree form of diazoxide (a) and the sodium salt of diazoxide (c). FTIRshowed expected changes for the sodium salt.

Elemental analysis of the salt indicated that the salt was formed in aratio of approximately 1:1, with the percentage of sodium being slightlylow (approximately 3.4%). This deficiency may be due to matrix effects,as NMR indicated the sample had a relatively high purity.

UV-vis measurements in neutral aqueous solution show a λmax ofapproximately 271 nm. (See FIG. 1). This value is slightly higher thanfree form diazoxide (265 nm). In acetonitrile, the λmax of the sodiumsalt exhibits a solvatochromic shift to approximately 296 nm. (See FIG.3). An increase in the pH of the solution is expected to produce abathochromic shift from approximately 265 nm to approximately 280 nm.

Solubility measurements performed at pH 2, 7, and 12 in 10 mM phosphatebuffer at room temperature showed solubility of the sodium salt ofdiazoxide to be 13.0 mg/mL, 18.1 mg/mL and 48.6 mg/mL, respectively.

Form conversion and degradation under thermal stress were conducted asdescribed for the free form diazoxide salt and showed the salt hadlittle propensity to change form or degrade over a period of 14 days.Similarly, slurry studies were conducted as described for the free formdiazoxide in n-heptane, dichloromethane and toluene showed no propensityof inter-conversion. See FIG. 6, wherein (a) is the XRPD pattern for thesodium salt of diazoxide, (b) is the XRPD pattern for the sodium saltafter the slurry study, and (c) the XRPD of the free form of diazoxide.

5.4. Characterization of Potassium Diazoxide Salt

The XRPD pattern for the potassium salt of diazoxide was analyzed,showing the material to be crystalline. (See FIG. 4( b)). The DSCanalysis revealed two major exothermic events at 128 and 354° C. (SeeFIG. 7). Small endotherms are likely due to sample impurities or thepresence of solvent. TGA analysis showed weight loss of 7.7% below 220°C., which may be the result of moisture sorption. (See FIG. 8).Theoretical weight loss for a monohydrate of the diazoxide salt is 6.6%.Moisture absorption performed from 0-90% relative humidity at 25° C.showed the material to be hygroscopic as the sample deliquesced at 90%relative humidity, showing 38.3 wt % water. The sample absorbed 5.4 wt %of water at 60% RH. Hysteresis was observed upon desorption, however thematerial was determined to be a hemihydrate from 0-30% RH and amonohydrate from 35-75% RH. XRPD following the desorption analysisindicated that the sample had changed to an alternate crystalline form.¹H NMR showed a chemical shift in the aromatic and methyl resonances ofthe sodium salt, as expected due to changes to the aromatic system. (SeeFIG. 5 showing spectrum of the free form diazoxide (a) and the Potassiumsalt of diazoxide (b)). FTIR showed expected changes for the potassiumsalt.

Elemental analysis of the potassium salt indicated that the salt wasformed in a ratio of approximately 1:1, with the percentage of potassiumbeing slightly low (approximately 1.6%). This deficiency may be due tomatrix effects, as NMR indicated the sample had a relatively highpurity.

UV-vis measurements of the potassium diazoxide salt in neutral aqueoussolution show a λ_(max) of approximately 265 nm, which is equivalent tothe diazoxide free form λ_(max). (See FIG. 1. In acetonitrile, theλ_(max) of the potassium salt exhibits a solvatochromic shift toapproximately 296 nm. (See FIG. 3). The potassium salt was used in a pHdependency study and showed that increasing the pH of the solutionresulted in a bathochromic shift of the λ_(max) from approximately 265nm to approximately 280 nm.

Solubility measurements performed at pH 2, 7, and 12 in 10 mM phosphatebuffer at room temperature showed solubility of the sodium salt ofdiazoxide to be 9.9 mg/mL, 14.4 mg/mL and 43.0 mg/mL, respectively. Thepotassium salt displayed greater solubility than the free formdiazoxide, and demonstrated similar solubility to the sodium diazoxidesalt. The XRPD pattern of solids obtained after the solubility analysisindicated that the potassium salt had changed back to the free formdiazoxide material.

Propensity for form conversion and degradation under thermal stress wereconducted as described for the free form diazoxide salt. The XRPDpattern of the sample after 7 and 14 days showed unique peaks, ascompared with the potassium salt starting material. Analysis by DSCafter 14 days also showed unique peaks as well. Using a gradient areapercent assay, HPLC did not show any significant degradation of thepotassium salt.

Slurry studies were conducted as described for the free form diazoxidein n-heptane, dichloromethane and toluene showed no propensity ofinter-conversion. XRPD analysis of samples after 7 and 14 days showedunique peaks similar to those observed with the thermal stress study.See FIG. 9, wherein (a) is the XRPD pattern of the potassium salt ofdiazoxide, (b) is the XRPD pattern of the potassium salt of diazoxideafter the slurry study in toluene, and (c) the XRPD pattern of thepotassium salt after 14 days of the slurry study in toluene. Analysis byDSC after 14 days also showed unique peaks as compared with the startingmaterial. HPLC using a gradient area percent assay did not show anysignificant degradation after the study.

5.5. Characterization of Choline Diazoxide Salt

The XRPD pattern of the choline salt of diazoxide was analyzed, showingthe material to be crystalline. (See FIG. 10 (b)). The DSC analysisrevealed a major exothermic events at 167° C. (See FIG. 11). A smallerendothermic event was seen at 119° C. and is likely due to sampleimpurities or the presence of residual solvent. TGA analysis showedweight loss of 0.8% between 100 and 140° C., which may be the result ofresidual solvents. (See FIG. 12). Moisture absorption performed from0-90% relative humidity at 25° C. showed the material to be hygroscopicas the sample absorbed over 28% at 80% relative humidity, anddeliquesced at 90% RH. Hysteresis was not observed upon desorption,indicating the choline salt does not form a stable hydrate. The XRPDpattern following the desorption analysis indicated that the sample hadchanged to an alternate crystalline form during the hydration andsubsequent desorption. (See FIG. 13 (c)) ¹H NMR was consistent with a1:1 molar ratio of diazoxide to counter-ion, and had the expecteddifferences in the chemical shift of the aromatic and methyl resonances,as expected due to the change in the local environment of the aromaticsystem due to the presence of the choline counter-ion. (See FIG. 14(b)). FTIR showed expected changes for the choline salt, similar tothose seen with the sodium and potassium salts.

Elemental analysis of the choline salt indicated that the salt wasformed in a ratio of approximately 1:1. This is consistent with the ¹HNMR.

UV-vis measurements of the choline diazoxide salt in neutral aqueoussolution show a λ_(max) of approximately 268 nm, which is close to theλ_(max) for the diazoxide free form of 265 nm. In acetonitrile, theλ_(max) of the choline salt exhibits a solvatochromic shift toapproximately 296 nm, which is consistent with the sodium and potassiumdiazoxide salts. The potassium salt was used in a pH dependency studyand showed that increasing the pH of the solution resulted in abathochromic shift of the λ_(max) from approximately 265 nm toapproximately 280 nm.

Solubility measurements performed at pH 2, 7, and 12 in 10 mM phosphatebuffer at room temperature showed solubility of the sodium salt ofdiazoxide to be 28.2 mg/mL, 41.5 mg/mL and greater than 293 mg/mL,respectively. The choline salt displayed greater solubility than thefree form diazoxide after being allowed to equilibrate for 12 hours. TheXRPD pattern of the solids obtained after the solubility analysis (at pH2 and 7 only) indicated that the choline salt had changed back to thefree form diazoxide material.

Propensity of the choline salt for form conversion and degradation underthermal stress were conducted as described for the free form diazoxidesalt. XRPD analysis of the sample after 7 and 14 days showed an XRPDpattern consistent with the starting material, as well as the presenceof additional unique peaks. (More unique peaks were present after 14days than after 7 days). Analysis by DSC after 14 days did not show anysignificant difference, with a small endotherm at 117° C. and a largeendotherm at 168° C. (Initial DSC showed endotherms at 119° C. and alarge endotherm at 167° C.). Using a gradient area percent assay, HPLCdid not show any significant degradation of the choline salt.

Slurry studies were conducted as described for the free form diazoxidein n-heptane, dichloromethane and toluene showed no propensity ofinter-conversion. XRPD analysis of samples after 7 and 14 days showedsignal associated with the starting material, with additional uniquepeaks present. (See FIG. 13 (c)). The XRPD pattern of slurry studysamples from n-heptane showed signals associated with the startingmaterial, as well as other additional unique signals. The XRPD patternof slurry samples from dichloromethane and toluene were consistent withspectra obtained after the thermal studies and the moisture sorptionanalysis. Analysis by DSC after 14 days revealed a small endotherm at109° C., and a major endotherm at 167° C. HPLC using a gradient areapercent assay did not show any significant degradation after the study.

5.6. Characterization of Diazoxide Salt of Hexamethyl HexamethyleneDiammonium Hydroxide (HHDADH)

The XRPD pattern of the HHDADH salt of diazoxide was analyzed, showingthe material to be a crystalline solid. (See FIG. 10( c)). Integrationof the ¹H NMR spectra was consistent with a 2:1 molar ratio of diazoxideto counter-ion (wherein the HHDADH counter-ion is divalent), and had theexpected differences in the chemical shift of the aromatic and methylresonances due to the change in the local environment of the aromaticsystem due to the presence of the choline counter-ion. (See FIG. 14(c)). The λ_(max) of the HHDADH diazoxide salt in acetonitrile measuredby UV-vis is 296 nm, which is consistent with the sodium, potassium andcholine diazoxide salts.

A summary of the characterization of the free form diazoxide and thepotassium, sodium, choline and hexamethyl hexamethylene diammoniumhydroxide salts of diazoxide are presented in Table 14.

TABLE 14 Summary of Characterization and Solubility for Diazoxide andSalts Potassium Sodium Choline Test Free form diazoxide salt diazoxidesalt diazoxide salt HHDADH XRPD crystalline crystalline crystallinecrystalline crystalline FTIR consistent consistent consistent consistentN/A DSC single multiple Single two N/A endotherm at endotherms belowexotherm endotherms 330° C. 300° C. above 400° C. below 200° C. UV in268 nm 265 nm 271 nm 268 nm N/A aqueous 264 nm 296 nm 296 nm 296 nm 296nm acetonitrile TGA <1% 7.7 (monohydrate <1% <1% N/A 6.7%) Moisture non-deliquescent - deliquescent - deliquescent - N/A Sorption hygroscopic -hemi/monohydrate potential no hysteresis no hysteresis hemihydrateSolubility pH 2.0 0.4 mg/mL 9.9 mg/mL 13.0 mg/mL 28.2 mg/mL N/A pH 7.00.04 mg/mL 14.4 mg/mL 18.1 mg/mL 41.5 mg/mL pH 12.0 4.8 mg/mL 43.0 mg/mL48.6 mg/mL >293 mg/mL

Additional solubility studies were conducted for diazoxide free baseform and diazoxide choline salt, as reported in Table 15. Eachdetermination was carried out in duplicate, slurrying each sample in a100 mM phosphate buffer solution, pH 7.00. Duplicate samples were thentitrated to pH 7.0 and pH 8.8 using a 0.1 N phosphoric acid solution,followed by stirring for 18 h at ambient temperature. After this timeall samples were centrifuged, and the supernatant was diluted withmobile phase (MeCN/Water.). Solubility was then obtained using acalibration curve for diazoxide by HPLC analysis. In Table 15, the term“Diazoxide, free from” refers to the free base of diazoxide; the term“Diazoxide, choline salt” is the choline salt of diazoxide as describedherein; the term “Diazoxide, choline salt, milled” refers to the cholinesalt of diazoxide which has been milled by methods described herein.Table 15 shows that in 100 mM phosphate buffer, pH 7, and with titrationwith 0.1N phosphoric acid to a pH of about 6.8 to 8.8, solubility isnotably suppressed compared to solubility at pH 10-11. Furthermore, thediazoxide choline salts were found to have increased solubility whencompared to the parent free base.

TABLE 15 Summary of Solubility for Diazoxide and Diazoxide Choline Salt.pH after Phosphate titration pH Sample buffer 1N with after amountamount H₃PO₄ 0.1N 18 h Solubility Sample (mg) (mL) (mL) H₃PO₄ slurry[mg/mL] Diazoxide, 49.23 1 0.01 6.87 6.90 0.07 free form Diazoxide 47.641 — 7.30 7.31 0.07 free form Diazoxide, 44.34 1 0.15 7.22 7.45 0.12choline salt Diazoxide, 48.26 1 0.11 8.81 8.64 0.18 choline saltDiazoxide, 45.35 1 0.14 7.32 7.36 0.12 choline salt, milled Diazoxide,53.15 1 0.16 8.50 8.52 0.20 choline salt, milled Diazoxide, 50.0 1 — —10.57 42.11 choline salt Diazoxide, 50.0 3 — — 10.52 32.88 choline salt,milled “—” Indicates no titration was conducted.

6. Polymorphic Forms of Diazoxide Salts

Polymorphic forms of the salts of diazoxide, characterization, andpreparation thereof are described.

6.1. Polymorphic Forms of the Choline Salt of Diazoxide

6.1.1. Demonstration of Preparation of Polymorphic Form B of the CholineSalt of Diazoxide

A 1-L round bottom flask was charged with diazoxide (5 g), MEK (750 mL)and choline hydroxide (5.25 g of 50 wt % solution in water), and heatedto 77° C. The mixture was allowed to cool to approximately ˜30° C., andfiltered to remove insoluble brown residues. The filtrate was thenconcentrated under reduced pressure to afford yellow oil, which wasdried in vacuo at 55° C. and 30 in. Hg to afford approximately 7.8 g asa waxy solid. The solids were dried in vacuo at 55° C. and 30 in. Hg toafford 7.13 g of the choline salt of diazoxide as a crystalline solid.Elemental analysis. Theoretical: C 46.77%, H 6.04% and N 12.59%.Measured: C 45.44%, H 5.98% and N 11.46%.

6.1.2. Characterization of the Polymorphic Form B of the Choline Salt ofDiazoxide

The polymorphic Form B of the choline salt of diazoxide was analyzed byXRPD, DSC, and ¹H NMR. As shown in FIG. 16, the two identifiedpolymorphic forms of the choline salt of diazoxide show different XRPDpatterns. See FIG. 16, wherein (a) shows the polymorphic Form A of thecholine salt of diazoxide.

As shown in FIG. 17, ¹H NMR of the polymorphic Form B of the cholinesalt of diazoxide shows no change from the polymorphic Form A.

Moisture absorption of the polymorphic Form B crystal structure of thecholine salt of diazoxide performed from 0-80% relative humidity (RH) at25° C. showed the material to be hygroscopic as the sample absorbed over14.5% at 70% relative humidity, and deliquesced at 80% RH. The XRPDpattern following the desorption analysis indicated that the sampleremained in the Form B crystal structure during the hydration andsubsequent desorption.

Solubility measurements performed at pH 2, 7, and 12 in 10 mM phosphatebuffer at room temperature showed solubility of the Form B crystalstructure of the choline salt of diazoxide to be 32.8 mg/mL, 80.1 mg/mLand 216 mg/mL, respectively. The XRPD pattern of the solids obtainedafter the solubility analysis (at pH 2 and 7) indicated that Form B ofthe choline salt of diazoxide was still present.

Slurry experiments were performed on each form to determine thepropensity for conversion and to search for possible new and/or uniqueforms. Upon slurrying Form B in CH2Cl2, n-heptane, and toluene, formconversion was not observed.

6.1.3. Demonstration of Preparation of the Polymorphic Form A of theCholine Salt of Diazoxide from the Polymorphic Form B of the CholineSalt of Diazoxide

Approximately 20 mg of the Form B polymorph of the choline salt ofdiazoxide was added to approximately 1 mL of acetone and heated toapproximately 55° C. The mixture was filtered while hot and placed in arefrigerator (4° C.) for 16 hours. No precipitate was observed. Thesolvent was evaporated down to dryness using a gentle stream ofnitrogen. The resultant solids were also dried in vacuo at roomtemperature and 30 in. Hg. XRPD analysis showed the salt had convertedfrom the Form B polymorph to the Form A polymorph.

6.1.4. Characterization of the Polymorphic Form A of the Choline Salt ofDiazoxide

Form A is an anhydrous crystalline form of diazoxide choline, with anendothermic event at approximately 165° C. in the DSC (see FIG. 20). TheXRPD pattern for Form A is unique compared to that of Form B as shown inFIG. 21. FTIR (ATR) spectroscopy additionally indicates differencesbetween the two forms. ¹H NMR analysis affords a spectrum consistentwith diazoxide and a 1:1 ratio of compound/counterion. NMR data alsoindicate that the magnetic environment of the diazoxide structurechanges between free and polymorph forms, as evidenced by a movement inchemical shift of the aromatic and methyl proton resonances. Inaddition, the resonance due to the amine proton is not observed whichsuggests deprotonation in solution. Weight loss by TGA is less than 1%and may be due to residual solvent. The temperature of weight loss isabove 100° C. which suggests that solvent may have been bound (i.e.,solvate material). Moisture-sorption analysis conducted at 25° C. from 0to 80% RH (adsorption) and 75 to 0% RH (desorption) shows Form A to be ahygroscopic solid, showing 2.4 wt % water at 60% RH. The sample wasfound to have deliquesced above 75% RH. In comparison, Form B is alsohygroscopic and showed 7.4 wt % water at 60% RH and deliquesced at 80%RH. XRPD analysis following the moisture-sorption experiment affords apattern consistent with Form A. Solubility studies conducted on bothforms at pH 2, 7, and 12 in phosphate buffer showed differences, withForm A showing 28, 41, and >293 mg/mL respectively. Solubilityconcentrations were determined using area-percent calculations with HPLCcalibration curves. Slurry experiments were performed on each form todetermine propensity for conversion and to see if a unique form could begenerated. Upon slurrying Form A in CH₂Cl₂, n-heptane, and toluene,conversion to Form B was observed after seven days. These resultssuggest that Form A is less thermodynamically stable under theseconditions than Form B according to Ostwald's Rule of Stages.

6.1.5. Screening for Polymorphic Forms of Diazoxide Choline Salt.

A polymorph screening study of diazoxide choline salt was conducted witha series of crystallization and slurry conditions. As described herein,interconversion of diazoxide choline salt forms A and B was observedduring this investigation. Each polymorphic form of diazoxide cholinesalt resulting from this study was characterized using the techniquesand procedures described herein. A summary of characterization tests islisted in Table 16.

TABLE 16 Characterization of Forms A and B of Diazoxide Choline Salt InScreening Study Experiment Form A Form B XRPD* Crystalline CrystallineDSC Endotherm ≈165° C. Endotherm ≈160° C. TGA <1% ≈1% FTIR (ATR)**Consistent w/structure Consistent w/structure ¹H NMR Consistent w/1:1ratio Consistent w/1:1 ratio Moisture Hygroscopic - deliquescedHygroscopic - deliquesced Sorption at 90% RH at 80% RH Solubility pH 2:28 mg/mL pH 2: 33 mg/mL pH 7: 41 mg/mL pH 7: 80 mg/mL pH 12: >293 mg/mLpH 12: 216 mg/mL Thermal Started to convert to Form Stable after 14 daysat B after 14 days at 60° C. 60° C. Slurries Converted to Form B afterStable after 14 days in 7 days in n-heptane, CH₂Cl₂, THF, and t-AmOHtoluene, and CH₂Cl₂ *Major peaks (2-θ): Form A (9.8, 10.5, 14.9, 17.8,17.9, 18.5, 19.5, 22.1, 22.6, 26.2, 29.6, 31.2); Form B (8.9, 10.3,12.0, 18.3, 20.6, 24.1, 24.5, 26.3, 27.1, 28.9). **Unique FTIR (ATR)absorbances (cm⁻¹): Form A (2926, 2654, 1592, 1449, 1248); Form B (3256,2174, 2890, 1605, 1463, 1235).

6.1.5.1. Solubility Screen in Organic Solvents.

Diazoxide choline, prepared in MEK using choline hydroxide as 50 wt %solution in water (see above) displayed some solubility in the followingsolvents: acetonitrile, acetone, ethanol, IPA, MEK, DMF, and methanol.These solvents were chosen due to differences in functionality,polarity, and boiling points and their ability to dissolve diazoxide.Other solvents which showed poor ability to dissolve salts were used asantisolvents and in slurry experiments where some solubility wasobserved: dioxane, MTBE, EtOAc, IPAc, THF, water, cyclohexane, heptane,CH₂Cl₂, and toluene.

Solvents for crystallizations during screening were chosen based on thesolubility screen summarized in Table 17. Crystallizations of diazoxidecholine from all conditions afforded a total of two forms, A and B.Forms A and B were found to be anhydrous polymorphs of diazoxidecholine. Form B was observed to be generated from most solvents used. Itwas difficult to isolate pure Form A on large scales (>50 mg) asconditions observed to produce Form A on a smaller scale (approximately50 mg or less) were found to result in Form B or mixtures of both formson larger scales. Based on room-temperature slurry experiments,anhydrous Form B was found to be the most thermodynamically stable formin this study. Form A readily converted to Form B in all slurry solventsutilized.

TABLE 17 Solubility Screen for Diazoxide Choline Salt Cmpd Solvent Conc.Temp. Solvent (mg) (mL) (mg/mL) (° C.) Soluble MeCN 1.7 0.25 >6.80 rtYes Dioxane 1.4 5.00 0.28 55 No Acetone 1.9 0.25 7.60 55 Yes MTBE 2.45.00 0.48 55 No EtOH 1.5 0.25 >6.00 rt Yes EtOAc 1.2 5.00 0.24 55 NoIPAc 1.4 5.00 0.28 55 No IPA 1.8 0.25 7.20 55 Yes THF 1.1 5.00 0.22 55No MEK 1.8 1.00 1.80 55 Yes DMF 1.2 0.25 >4.80 rt Yes Water 2.0 5.000.40 55 No MeOH 1.9 0.25 7.60 55 Yes c-Hexane 2.0 5.00 0.40 55 NoHeptane 1.9 5.00 0.38 55 No CH₂Cl₂ 1.3 5.00 0.26 55 Partially Toluene1.4 5.00 0.28 55 No

6.1.5.2. Single-Solvent Crystallizations

Fast cooling procedure: Diazoxide (approximately 20 mg) was weighed outinto vials and enough solvent (starting with 0.25 mL) was added untilthe material completely dissolved at elevated temperature. After hotfiltration the vials were placed in a refrigerator (4° C.) for 16 hours.After the cooling-process the samples were observed for precipitateswhich were isolated by filtration. Vials not demonstrating precipitateswere evaporated down to dryness using a gentle stream of nitrogen. Allsolids were dried in vacuo at ambient temperature and 30 in. Hg.

Slow cooling procedure: Diazoxide (approximately 30 mg of choline salt)was weighed out into vials and enough solvent was added until thematerial went into solution at elevated temperature. After hotfiltration the vials were then slowly cooled to room temperature at therate of 20° C./h and stirred at room temperature for 1-2 hours. Allsolids were dried in vacuo at ambient temperature and 30 in. Hg.

Based on the initial solubility study, seven solvents were selected forthe fast-cooling crystallization: acetonitrile, acetone, ethanol, IPA,MEK, DMF, and methanol. Table 18 shows a list of the solvents that wereused and the amount of solvent needed to dissolve the material. Afterthe cooling-process precipitates were noticed in samples #2, 3, 5, and6, the solids were isolated by filtration. The other samples (#1, 4, and7) were evaporated down to dryness using a gentle stream of nitrogen.The diazoxide choline salts were found to be consistent with Form A byXRPD analysis for all solids with the exception of sample #2 (consistentwith the freeform) and sample #5 (consistent with Form B with preferredorientation observed).

TABLE 18 Single-Solvent Crystallization of Diazoxide Choline Salt UsingFast-Cooling Procedure BP Cmpd Solvent Amt Conc Temp. Entry Solvent (°C.) (mg) (mL) (mg/mL) (° C.) Precipitate Form 1 Acetone 56 21.0 1.0021.00 55 No/Evap A 2 MeOH 64 20.3 0.25 81.20 55 Yes FF* 3 EtOH 78 21.30.25 85.20 62 Yes A 4 MEK 80 19.6 1.25 15.68 75 No/Evap A 5 MeCN 81 20.60.25 82.40 55 Yes Unique 6 IPA 82 22.8 0.25 91.20 62 Yes A 7 DMF 15326.0 0.25 104.00 55 No/Evap A

In accordance with the data obtained from fast-cooling experiments, foursolvents which showed precipitation of solids were chosen for theslow-cooling experiments: MeOH, EtOH, MeCN, and IPA (Table 19). Allobtained analyzable solids of the choline salt were found to beconsistent with Form B by XRPD with the exception of Entry #1 which wasconsistent with diazoxide freeform and Entry #2 which was notanalyzable. Mother liquor of Entry #2 was concentrated to dryness andthe residual solids were analyzed by XRPD and found to be Form Bmaterial. As a result of obtaining freeform material from thesingle-solvent crystallizations in methanol, three more alcohols weretested for the single-solvent crystallizations using fast- andslow-cooling procedures. Tables 20 and 21 provide a list of the solventsthat were used and the amount of solvent needed to dissolve thematerial. XRPD patterns of the fast-cooling procedure showed freeform ofdiazoxide from isobutanol, Form B from isoamyl alcohol, and Form A fromtert-amyl alcohol compared to the slow-cooling procedure, which affordedForm B material from all three solvents.

TABLE 19 Single-Solvent Crystallization of Diazoxide Choline Salt UsingSlow-Cooling Procedure Boiling Material Solvent Conc. Pre- Point AmountAmount (mg/ Temp. cip- Solvent (° C.) (mg) (mL) mL) (° C.) itate FormMeOH 64 32.1 0.3 107.00 62 Yes FF* EtOH 78 33.3 0.3 111.00 75 Yes NA**MeCN 81 30.9 0.3 103.00 62 Yes B IPA 82 33.7 0.3 112.33 80 Yes B

TABLE 20 Single-Solvent Crystallization of Diazoxide Choline Salt UsingFast-Cooling Procedure Boiling Material Solvent Conc. Point AmountAmount (mg/ Temp. Precip- Solvent (° C.) (mg) (mL) mL) (° C.) itate Formi-BuOH 108 29.7 0.3 99.00 78 Yes (sm)* i-AmOH 130 29.6 0.3 98.67 82 YesB t-AmOH 102 29.5 0.3 98.33 95 No/ A Evap

TABLE 21 Single-Solvent Crystallization of Diazoxide Choline Salt UsingSlow-Cooling Procedure Boiling Material Solvent Conc. Pre- Point AmountAmount mg/ Temp. cip- Solvent (° C.) (mg) (mL) (mL) (° C.) itate Formi-BuOH 108 33.0 0.3 110.00 92 Yes B i-AmOH 130 28.2 0.3 94.00 92 Yes Bt-AmOH 102 29.0 0.4 72.50 92 Yes B

The results of the choline salt single-solvent fast- and slow-coolingcrystallizations (see Tables 19 to 21) indicated that Form A was morelikely to be isolated with fast-cooling profiles and Form B withslow-cooling profiles.

6.1.5.3. Binary Solvent Crystallizations

Binary-solvent crystallizations of the choline salt were performed usingfour primary solvents (MeOH, EtOH, IPA, and MeCN) and nine cosolvents(MTBE, EtOAc, IPAc, THF, c-hexane, heptane, toluene, CH₂Cl₂, anddioxane) with a fast-cooling profile (supra). XRPD patterns showed thatForm B was obtained from mixtures of MeOH with MTBE, EtOAc, IPAc,toluene, and dioxane. As shown in Table 22, Form A was obtained frommixtures of MeOH with THF and with CH₂Cl₂ after evaporating the solventto dryness. The mixtures of MeOH with cyclohexane and heptane providedthe freeform of diazoxide. All solids obtained from fast-coolingprocedures with EtOH, IPA, and MeCN as primary solvents provided Form Bmaterial.

TABLE 22 Binary-Solvent Crystallizations of Choline Salt of DiazoxideUsing Fast-Cooling Procedure and MeOH as a Primary Solvent DiazoxideMeOH* Amount Amt (mg) (mL) Antisolvent (mL) Precipitate Form 27.8 0.3MTBE 1.4 Yes B 30.7 0.3 EtOAc 6.0 Yes B 32.0 0.3 IPAc 6.0 Yes B 31.9 0.3THF 6.0 No/Evap to Dry A 29.5 0.3 c-Hexane 2.0 Yes (small) FF** 30.2 0.3Heptane 2.0 Yes FF** 29.3 0.3 Toluene 6.0 Yes B 32.0 0.3 CH₂Cl₂ 6.0No/Evap to Dry A 28.8 0.3 Dioxane 6.0 Yes B *Solids were dissolved at62° C. **Freeform of diazoxide.

Binary-solvent recrystallizations of the choline salt with theslow-cooling procedure were performed using two primary solvents (IPAand MeCN) and nine cosolvents (MTBE, EtOAc, IPAc, THF, c-hexane,heptane, toluene, CH₂Cl₂, and dioxane). All solids obtained from aslow-cooling procedure with IPA and MeCN as primary solvents providedForm B material based on XRPD analysis. The results of binary-solventcrystallizations indicated that Form B was the most thermodynamicallystable form of diazoxide choline.

6.1.5.4. Binary Solvent Crystallizations Using Water as a Cosolvent

In an attempt to investigate the formation of hydrates of the cholinesalt, experiments was performed using fast- and slow-cooling proceduresand water as a cosolvent.

The fast cooling procedure (supra) was used with the exception of usingdifferent primary solvents which were miscible with water: acetone,acetonitrile, DMF, IPA, i-BuOH, i-AmOH, and t-AmOH. Water was utilizedin these crystallizations as a cosolvent. All solids obtained from thefast-cooling procedure with water as the cosolvent provided diazoxidefreeform material by XRPD analysis.

To compare the results obtained from the fast-cooling procedure a set ofexperiments was performed using a slow-cooling procedure and water as acosolvent. All obtained solids were analyzed by XRPD and affordedpatterns consistent with diazoxide freeform. Without wishing to be boundby theory, these results suggest that the conditions used forcrystallization caused dissociation of the choline salt. A small amountof a second crop was obtained in each sample, but only two samples wereanalyzable by XRPD and indicated that the samples were freeformmaterial. All mother liquors were evaporated to dryness and the residualsolids were also analyzed by XRPD to afford patterns consistent withForm B of the choline salt.

6.1.5.5. Metastable Zone Width Estimation

Form B: To produce a robust process, an understanding of the solubilityprofiles of the various solid forms under consideration is required.From a practical standpoint, this involves the measurement of themetastable zone width (MSZW) of pure forms, whereby the saturation andsupersaturation curves of the different forms are generated over a welldefined concentration and temperature range. This knowledge can then beused to design a crystallization protocol that should ideally favor aselective crystal growth of the desired form.

Form B of diazoxide choline salt showed moderate solubility in a solventmixture made of MeCN/MeOH/MtBE (10:1:12, volume ratios). The wide widthof the metastable zone as shown in Table 23 gives many seeding options.During the MSZW measurement, aliquots from the crystallizing materialwere withdrawn and analyzed by XRPD to ensure that no form conversionoccurred during the experiment. Indeed, the material remained unchangedduring the test.

TABLE 23 Meta-Stable Zone Width For Form B Diazoxide Choline Salt inMeCN/MeOH/MtBE (10:1:12) (v/v). Conc.| Temp. In Temp. Out Temp. Range(mg/mL) (° C.) (° C.) (° C.) 30.8 53.2 35.0 18.2 28.5 49.0 33.6 15.426.5 47.0 32.0 15.0 24.7 43.8 29.1 14.7 23.2 40.5 28.5 12.0 21.9 38.026.0 12.0

Form A: The metastable zone width for Form could not be estimatedbecause this polymorphic form converted during the experiment to Form B.

6.1.5.6. Crystallization of Form A of Diazoxide Choline Salt

The choline salt of diazoxide (160.3 mg) was dissolved in 1 mL of IPA at55° C. which was then passed through a Millipore 0.45 μM filter into aclean vial. This vial was placed in freezer a −20° C. overnight. Solidswere not noticed and the flask was scratched with a micro-spatula. Thevial was placed back in the freezer and nucleation was noticed after tenminutes. The solids were collected by vacuum filtration and washed with1 mL of MtBE. The solids were dried in vacuo at 40° C. and 30 in. Hg toafford 70 mg (43.6% recovery) of Form A as determined by XRPD.

6.1.5.7. 500-mg Scale Crystallization of Form B of Diazoxide CholineSalt

The choline salt of diazoxide (524.3 mg) was dissolved in 3 mL of IPA at78° C. and this solution was then cooled to 55° C. for the addition ofMtBE. The MtBE (4 mL) was added until nucleation was observed. Afternucleation the batch was allowed to cool to room temperature at a rateof 20° C./h. The solids were collected by vacuum filtration and washedwith 1 mL of MtBE. The solids were dried in vacuo at 40° C. and 30 in.of Hg to afford 426.7 mg (81.3% recovery) of Form B as determined byXRPD.

6.1.5.8. 2-g Scale Crystallization of Form B of Diazoxide Choline Salt

The choline salt of diazoxide (2.0015 g) was dissolved in 5.5 mL of IPAat 78° C. to afford a clear solution. This solution was passed through aMillipore Millex FH 0.45 μM filter. This solution was then cooled to 55°C. MtBE was added in 1 mL portions, with a two minute interval betweenportions. Nucleation was noted after the second addition of MtBE. Thissuspension was allowed to cool to room temperature at a rate of 20° C./hand stirred at this temperature for 16 hours. The solids were collectedby vacuum filtration and washed with 1 mL of MtBE. The solids were driedin vacuo at 40° C. and 30 in. of Hg to afford 1.6091 g (80.4% recovery)of Form B as determined by XRPD.

6.1.5.9. Detection of Form Impurities

Mixtures of diazoxide choline Forms A and B were prepared by adding aminor amount of Form A to Form B. Samples were lightly ground by handswith a mortar and pestle for approximately one minute. Samples were thenanalyzed by XRPD analysis. XRPD analysis was found to be suitable fordetecting 5% of Form A in Form B.

6.2. Polymorphic Forms of the Potassium Salt of Diazoxide

A summary of characterization tests for three common crystalline formsof diazoxide potassium salt are listed in Table 24. Solvents forcrystallizations were chosen based on the solubility screen summarizedbelow. Crystallizations of diazoxide potassium salt from all conditionsprovided herein afforded a total of seven unique crystalline forms, Athrough G. Forms C, D, and F were found to be the most common during thecrystallization screen, and were therefore scaled up for furthercharacterization.

TABLE 24 Results Summary of Characterization Tests for Salt of DiazoxidePotassium Experiment Form C Form D Form F XRPD* Crystalline CrystallineCrystalline DSC 187, 360° C. 130, 191, 352° C. 191, 363° C. TGA 8.4%**4.5%** 13.1%** FTIR (ATR)* Consistent w/ Consistent w/ Consistent w/structure structure structure ¹H NMR Consistent w/ Consistent w/Consistent w/ structure** structure structure** Moisture Hygroscopic -Hygroscopic - Sorption deliquesced at 90% RH deliquesced at 90% RHHygroscopic - deliquesced at 90% RH Solubility N/A pH 2: 29 mg/mL N/A pH7: 33 mg/mL pH 12: 59 mg/mL Thermal Stable (7 days) Stable (7 days)Stable (7 days) Slurries Converted to Form D Converted to Form DConverted to Form D *XRPD Major peaks (2-θ): Form A (6.0, 8.1, 16.3,17.7, 18.6, 19.1, 22.9, 23.3, 23.7, 24.7, 25.4, 26.1, 28.2, 29.6, 30.2);Form B (8.5, 10.8, 16.9, 18.2, 21.6, 25.5, 26.1, 28.9); Form C (5.7,6.1, 17.9, 23.9, 25.1, 37.3); Form D (5.7, 6.2, 8.1, 8.5, 8.8, 16.9,18.6, 23.2, 24.5, 25.8, 26.1); Form E (6.7, 7.1, 14.1, 21.2); Form F(8.5, 9.0, 18.7, 20.6, 23.5, 27.5, 36.3); Form G (5.2, 5.5, 13.1, 16.5,19.3, 22.8, 24.8, 26.4, 28.7, 3 4.1); *Unique FTIR (ATR) absorbances(cm⁻¹): Form A (1503, 1374, 1339, 1207, 1131, 1056, 771); Form B (1509,1464, 1378, 1347); Form C (1706, 1208, 1146, 746); Form D (1595, 1258,1219, 890); Form E (1550, 1508, 1268, 1101, 1006). Form F (1643, 1595,1234, 1145, 810). Form G (1675, 1591, 1504, 1458, 1432, 1266, 999, 958,905, 872). **Data indicates a half-molar equivalent of acetone, water,or dioxane for Forms C, D, and F respectively.

Diazoxide potassium Forms C, D, and F were observed to be an acetonesolvate, a hemihydrate, and a dioxane solvate of diazoxide potassium,respectively. Form C is an acetone solvate that was generatedpredominantly when acetone was used in the crystallization. Form D, ahemihydrate, was observed to be generated from most solvents used. FormF is a dioxane solvate generated when dioxane was used as anantisolvent. Forms A, B, E, and G were not commonly observed during thecrystallization. Elemental analysis data indicated that the unique formsobserved may be mixtures and/or have residual solvent(s) present.

Based on room-temperature slurry experiments presented, Form D was foundto be the most thermodynamically stable form of those discovered in thisstudy. Forms C and F readily converted to Form D in all slurry solventsutilized. Without wishing to be bound by theory, since nonaqueoussolvents were used, the material may have converted to the hemihydrate,Form D, upon removal from the solvent.

6.2.1. Demonstration of Preparation of Polymorphic Form A of thePotassium Salt of Diazoxide

The polymorphic Form A of the potassium salt of diazoxide was preparedas described above.

6.2.2. Demonstration of Preparation of the Polymorphic Form B of thePotassium Salt of Diazoxide

Diazoxide (2.95 g) was combined with 450 mL of methyl ethyl ketone andheated to approximately 77° C. to dissolve the diazoxide. To thesolution was added approximately 13 mL of 1M potassium hydroxide at arate of approximately 20 mL/min, stirred and allowed to cool to roomtemperature. The solution was stirred at room temperature for ˜16 h. Thesolvent was removed under reduced pressure, and the residual solids weredried in vacuo at 57° C. and 30 in. Hg to afford 3.7 g of the potassiumsalt.

6.2.3. Characterization of the Polymorphic Form B of the Potassium Saltof Diazoxide

The Form B polymorph of the potassium salt of diazoxide was analyzed byXRPD, and ¹H NMR. FIG. 18 shows the XRPD pattern of (a) the Form Apolymorph of the potassium salt of diazoxide and (b) the Form Bpolymorph of the potassium salt of diazoxide.

The ¹H NMR of the Form B polymorph of the potassium salt of diazoxideshows no change from the Form A polymorph.

6.2.4. Demonstration of Preparation of the Polymorphic Form A of thePotassium Salt of Diazoxide from Form B

Approximately 20 mg of the Form B polymorph of the diazoxide potassiumsalt was added to 2 mL of acetone and heated until the materialcompletely dissolved at 55° C. The solution was hot filtered and placedin a refrigerator (4° C.) for 16 hours. No precipitate was formed. Thesolvent was evaporated down to dryness using a gentle stream of nitrogenand the resultant solids were dried in vacuo at room temperature and 30in. Hg. The solid was analyzed by XRPD to determine the physical form.See FIG. 18( a).

6.2.5. Preparation of the Polymorphic Form C of the Potassium Salt ofDiazoxide from Form B

Approximately 20 mg of the Form B polymorph of the diazoxide potassiumsalt was added to 6 mL of ethyl acetate and heated until the materialcompletely dissolved at 75° C. The solution was hot filtered and placedin a refrigerator (4° C.) for 16 hours. No precipitate was formed. Thesolvent was evaporated down to dryness using a gentle stream of nitrogenand the resultant solids were dried in vacuo at room temperature and 30in. Hg. The solid was analyzed by XRPD to determine the physical form.See FIG. 18( c).

6.2.6. Preparation of the Polymorphic Form D of the Potassium Salt ofDiazoxide from Form B

Approximately 20 mg of the Form B polymorph of the diazoxide potassiumsalt was added to 0.3 mL of isopropyl alcohol and heated until thematerial completely dissolved at 62° C. The solution was hot filteredand placed in a refrigerator (4° C.) for 16 hours. No precipitate wasformed. The solvent was evaporated down to dryness using a gentle streamof nitrogen and the resultant solids were dried in vacuo at roomtemperature and 30 in. Hg. The solid was analyzed by XRPD to determinethe physical form. See FIG. 19( a).

6.2.7. Preparation of the Polymorphic Form E of the Potassium Salt ofDiazoxide from Form B

Approximately 20 mg of the Form B polymorph of the diazoxide potassiumsalt was added to 2.5 mL of tert-amyl alcohol and heated until thematerial completely dissolved at 95° C. The solution was hot filteredand placed in a refrigerator (4° C.) for 16 hours. No precipitate wasformed. The solvent was evaporated down to dryness using a gentle streamof nitrogen and the resultant solids were dried in vacuo at roomtemperature and 30 in. Hg. The solid was analyzed by XRPD to determinethe physical form. See FIG. 19(B).

6.2.8. Preparation of the Polymorphic Form F of the Potassium Salt ofDiazoxide from Form B

Approximately 20 mg of the Form B polymorph of the diazoxide potassiumsalt was added to 0.6 mL of acetonitrile and heated until the materialcompletely dissolved at 80° C. The solution was hot filtered, 6 mL ofdioxane was added, and the solution was placed in a refrigerator (4° C.)for 16 hours. No precipitate was formed. The solvent was evaporated downto dryness using a gentle stream of nitrogen and the resultant solidswere dried in vacuo at room temperature and 30 in. Hg. The solid wasanalyzed by XRPD to determine the physical form. See FIG. 19(C).

6.2.9. Preparation of the Polymorphic Form G of the Potassium Salt ofDiazoxide from Form B

Approximately 22.3 mg of the Form B polymorph of the diazoxide potassiumsalt was added to 0.5 mL of isoamyl alcohol and heated until thematerial completely dissolved at 73° C. The solution was hot filtered, 6mL of isopropyl acetate was added, and the solution was placed in arefrigerator (4° C.) for 16 hours. No precipitate was formed. Thesolvent was evaporated down to dryness using a gentle stream of nitrogenand the resultant solids were dried in vacuo at room temperature and 30in. Hg. The solid was analyzed by XRPD to determine the physical form.See FIG. 19(D).

As shown in Tables 25 and 26, both the solvent used forrecrystallization and the rate of cooling (i.e., fast cooling vs. slowcooling during the recrystallization) effect the crystal structureobtained once the product is isolated.

TABLE 25 Fast cooling of Potassium Salt of Diazoxide in Various SolventsCmpd Solvent Recovery Solvent (mg) (mL) (mg) Form THF 20.8 6.5 14.2 BEtOAc 21.1 6.0 10.6 C MeCN 21.2 0.5 n/a C + D IPA 22.7 0.3 n/a D Water20.6 1.5 n/a Free form diazoxide tAA 21.9 2.5 16.7 E IAA 19.7 0.3 n/a DDMF 21.6 0.3 15.9 G

TABLE 26 Slow cooling of Potassium Salt of Diazoxide in Various SolventsCmpd Solvent Recovery Solvent (mg) (mL) (mg) Form EtOAc 20.4 6.0 7.1 DMeCN 22.8 0.5 13.8 C IPA 21.5 0.3 13.2 C Water 20.9 1.5 1.2 Free formdiazoxide tAA 21.5 2.5 15.4 E IAA 20.6 0.3 6.6 D

6.2.10. Polymorphs Obtained by Recrystallization of the Potassium Saltof Diazoxide in Binary Solvents

As shown in Tables 27 and 28, recrystallization of the potassium salt ofdiazoxide from a variety of binary solvent systems also demonstratedconversion of the potassium salt to an alternate form. Use ofacetonitrile as the primary solvent is shown in Table 27 and use ofacetone as the primary solvent is shown in Table 28. As shown in Table27, recrystallization of the Form B polymorph of the potassium salt ofdiazoxide from acetonitrile using methyl tert-butyl ether, ethylacetate, isopropyl acetate, tetrahydrofuran, c-hexane, heptane, tolueneand dichloromethane as the secondary solvent all yielded the D Formpolymorph of the potassium salt. Recrystallization from acetonitrileusing dioxane as the secondary solvent yielded the F Form polymorph ofthe diazoxide salt of potassium.

TABLE 27 Recrystallization in Acetonitrile Acetonitrile Secondary CmpdRecovery (mL) solvent (mg) (mg) Form 0.6 MTBE 20.7 13.7 D 0.6 EtOAc 23.49.5 D 0.6 IPAc 20.0 13.3 D 0.6 THF 20.3 6.4 D 0.6 c-Hexane 20.4 9.6 D0.6 Heptane 20.3 10.8 D 0.6 Toluene 23.6 16.1 D 0.6 Dichloromethane 21.312.7 D 0.6 Dioxane 20.7 12.6 F

As shown in Table 28, recrystallization of the Form B polymorph of thepotassium salt of diazoxide from acetone using methyl tert-butyl ether,tetrahydrofuran, and c-hexane as the secondary solvent all yielded theForm A polymorph of the potassium salt of diazoxide. Recrystallizationfrom acetone using ethyl acetate, heptane, toluene and dichloromethaneas the secondary solvent all yielded the C Form polymorph of thepotassium salt. Recrystallization from acetone using isopropyl acetateas the secondary solvent yielded the D Form polymorph of the diazoxidesalt of potassium. Recrystallization from acetone using dioxane as thesecondary solvent yielded the F Form polymorph of the diazoxide salt ofpotassium.

TABLE 28 Recrystallization in Acetone Acetone Amt Recovery (mL) OtherSolvent (mg) (mg) Form 2.0 MTBE 20.2 13.9 A 2.0 EtOAc 21.6 4.8 C 2.0IPAc 20.6 11.6 D 2.0 THF 20.9 12.0 A 2.0 c-Hexane 21.3 12.3 A 2.0Heptane 20.6 12.7 C 2.0 Toluene 20.4 13.3 C 2.0 Dichloromethane 21.113.0 C 2.0 Dioxane 20.4 12.5 F

6.2.11. Screening for Polymorphic Forms of Diazoxide Potassium Salt.

A polymorphic screening study of diazoxide potassium salt was conductedwith a series of crystallization conditions described below.

6.2.11.1. Solubility Screening for Polymorphic Forms of DiazoxidePotassium Salt.

Diazoxide potassium, prepared in MEK using 1 M potassium hydroxidesolution in water, displayed some solubility in the following tensolvents: acetone, THF, EtOAc, MEK, MeCN, IPA, water, t-AmOH, i-AmOH,and DMF. These solvents were chosen due to differences in functionality,polarity, and boiling points and their ability to dissolve diazoxide.Solvents affording poor to fair solubility were used as antisolvents inbinary/ternary crystallizations as well as slurry studies. Table 29summarizes the results of the solubility screen.

TABLE 29 Solubility of Diazoxide Potassium Salt in Various Solvent CmpdAmt Solvent Conc. Temp. Solvent (mg) (mL) (mg/mL) (° C.) Soluble MeCN1.7 2.00 0.85 55 Yes Dioxane 1.4 5.00 0.28 55 No Acetone 1.6 4.00 0.4055 Yes MTBE 1.8 5.00 0.36 55 No EtOH 2.2 0.75 2.93 55 Yes EtOAc 1.8 5.000.36 55 No IPAc 1.7 5.00 0.34 55 No IPA 2.1 1.00 2.10 55 Yes THF 1.85.00 0.36 55 Partially MEK 1.5 5.00 0.30 55 Partially DMF 1.6 0.25 >6.40rt Yes Water 1.5 5.00 0.30 55 No MeOH 1.5 0.25 6.00 55 Yes c-Hexane 1.55.00 0.30 55 No Heptane 1.2 5.00 0.24 55 No CH₂Cl₂ 1.3 5.00 0.26 55 NoToluene 1.4 5.00 0.28 55 No

6.2.11.2. Single-Solvent Screening for Polymorphic Forms of DiazoxidePotassium Salt.

Single-solvent crystallizations of potassium salt were performed usingten solvents: acetone, THF, EtOAc, MEK, MeCN, IPA, water, t-AmOH,i-AmOH, and DMF for the fast-cooling procedure and six solvents (EtOAc,MeCN, IPA, water, t-AmOH, and i-AmOH) for the slow-cooling procedures.The “fast” and “slow” cooling procedures were as described above. Fourof the solvents were excluded from the slow-cooling experiments becausethey did not provide solids during fast-cooling experiments and neededto be evaporated to dryness. Tables 30 and 31 provide a list of thesolvents that were used and the amount of solvent needed to dissolve thematerial. All solids were analyzed by XRPD to determine the physicalform and six unique patterns (Forms A-E, G) were observed.

TABLE 30 Single-Solvent Crystallizations of Potassium Salt of DiazoxideUsing Fast-Cooling Cmpd Solvent Conc BP Amt Amt (mg/ Temp Solvent (° C.)(mg) (mL) mL) (° C.) Precipitate Form Acetone 56 20.7 2.0 10.35 55No/Evap A THF* 65 20.8 6.5 3.20 63 No/Evap B EtOAc 76 21.1 6.0 3.52 75Yes C MEK 80 20.2 4.0 5.05 75 No/Evap A MeCN 81 21.2 0.5 42.40 80 YesC + D IPA 82 22.7 0.3 75.67 62 Yes D Water 100 20.6 1.5 13.73 95 Yes FFt-AmOH 103 21.9 2.5 8.76 95 Yes E i-AmOH 130 19.7 0.3 65.67 73 Yes D DMF153 21.6 0.3 72.00 RT No/Evap G *Solids were not completely dissolved.

TABLE 31 Single-Solvent Crystallizations of Potassium Salt of DiazoxideUsing Slow- Cmpd Solvent Conc BP Amt Amt (mg/ Temp Solvent (° C.) (mg)(mL) mL) (° C.) Precipitate Form EtOAc 76 20.4 6.0 3.40 75 Yes D MeCN 8122.8 0.5 45.60 80 Yes C IPA 82 21.5 0.3 71.67 80 No/Evap C Water 10020.9 1.5 13.93 95 Yes FF t-AmOH 103 21.5 2.5 8.60 95 Yes E i-AmOH 13020.6 0.3 68.67 80 Yes D

6.2.11.3. Binary-Solvent Screening for Polymorphic Forms of DiazoxidePotassium Salt.

Binary-solvent crystallizations of the potassium salt utilizingfast-cooling procedure were performed using MeCN, acetone, and isoamylalcohol as primary solvents and the following nine cosolvents: MTBE,EtOAc, IPAc, THF, c-hexane, heptane, toluene, CH₂Cl₂, and dioxane. Table32 is representative, employing acetonitrile as primary solvent. XRPDpatterns from crystallizations using acetonitrile as a primary solventwere consistent with Form D with only one exception being the solidsobtained from the mixture of MeCN/dioxane afforded a unique pattern(Form F) material.

TABLE 32 Binary-Solvent Crystallizations of Potassium Salt of DiazoxideUsing Fast-cooling Procedure and McCN as a Primary Solvent Cmpd AmtMcCN* Anti- Amt (mg) (mL) solvent (mL) Precipitate Form 20.7 0.6 MTBE1.0 Yes D 23.4 0.6 EtOAc 6.0 Yes D 20.0 0.6 IPAc 3.0 Yes D 20.3 0.6 THF6.0 No/Evap. D To ppt 20.4 0.6 c-Hexane 2.0 Yes D 20.3 0.6 Heptane 2.0Yes D 23.6 0.6 Toluene 1.0 Yes D 21.3 0.6 CH₂Cl₂ 1.0 Yes D 20.7 0.6Dioxane 6.0 Yes F *Solids were dissolved at 80° C.

XRPD patterns from binary crystallizations with acetone as primarysolvent employing a fast cooling procedure afforded four forms A, C, D,and F. Mixtures of acetone with MTBE, THF, and cyclohexane provided FormA material; Form C was obtained from the mixtures of acetone with EtOAc,heptane, toluene, and CH₂Cl₂. The mixture of acetone with IPAc affordedForm D, and the mixture of acetone with dioxane afforded Form F solids.

XRPD patterns from binary crystallizations with isoamyl alcohol as theprimary solvent employing a fast cooling procedure afforded five forms:C, D, E, F, and G. Form C was obtained from crystallizations using MTBEand EtOAc as cosolvents; Form D was obtained from mixtures of isoamylalcohol with heptane, toluene, and CH₂Cl₂; Form E was crystallized outof i-AmOH/THF and i-AmOH/cyclohexane; Form G was obtained fromi-AmOH/IPAc and Form F was obtained from i-AmOH/dioxane. Form D was themost common form observed from the crystallizations, and Form F wasobserved only when dioxane was used as an antisolvent.

Binary solvent recrystallizations of the potassium salt with theslow-cooling procedure were performed using three primary solvents(MeCN, acetone, and i-AmOH) and eight cosolvents (MTBE, EtOAc, IPAc,c-hexane, heptane, toluene, CH₂Cl₂, and dioxane). All solids wereanalyzed by XRPD to determine the physical form. Two patterns wereobserved to be Forms C and D respectively with additional peaks present.Other crystallizations provided Forms D, C, or F. Form D was obtainedfrom the following solvent mixtures: MeCN/MTBE, MeCN/IPAc, MeCN/toluene,MeCN/CH₂Cl₂ and also from the mixtures of i-AmOH with MTBE, IPAc,cyclohexane, heptane, and toluene. Form C was obtained fromMeCN/heptane, i-AmOH/EtOAc, and mixtures of acetone with MTBE, EtOAc,IPAc, cyclohexane, heptane, toluene, and CH₂Cl₂. Form F was crystallizedfrom the mixtures of MeCN/dioxane and i-AmOH/dioxane. The solventmixture of MeCN/EtOH provided amorphous material. Elemental analysisresults indicate that the forms observed may not be pure and/or havebound or residual solvents present. Forms C, D, and F were found to bethe most common forms of the potassium salt isolated and based on theresults, these forms were chosen for scale-up and furthercharacterization. Differences were found in the XRPD patterns and FTIRspectra of the scale-up lots which were attributed to differences inimpurity profiles, crystallinity, and form purity.

6.2.11.4. Characterization of Polymorphic Form C of Diazoxide PotassiumSalt.

Form C of diazoxide potassium salt is an acetone solvate with a 2:1ratio of diazoxide/solvent. It is a crystalline form of diazoxidepotassium, with endothermic events at 187 and 360° C. in the DSC. TheXRPD pattern for Form C is unique compared to all other forms observed.FTIR (ATR) spectroscopy showed differences between forms. ¹H NMR spectrawere found to be consistent with the structure of diazoxide with ahalf-molar equivalent of acetone present. NMR data also indicated thatthe magnetic environment of the diazoxide structure had changedevidenced by a movement in chemical shift of the aromatic and methylproton resonances. In addition, the resonance due to the amine protonwas not observed which suggested deprotonation in solution. Weight lossby TGA was 8.9%, consistent with a half-molar equivalent of acetone, andoccurred near 180° C. consistent with the endotherm observed in the DSCexperiment. Moisture-sorption analysis conducted at 25° C. from 0 to 90%RH (adsorption) and 85 to 0% RH (desorption) showed Form C to be ahygroscopic solid, showing approximately 47 wt % water at 90% RH whichindicated the sample deliquesced. In comparison, Forms D and F(hemihydrate and dioxane solvate) showed approximately 26 and 22 wt %water at 90% RH respectively. XRPD analysis following themoisture-sorption experiment afforded a pattern consistent with Form D.Slurry experiments were performed on 50/50 mixtures of Forms C, D, and Fto determine propensity for conversion and to see if a unique form couldbe generated. Upon slurrying Form C mixtures in ethyl acetate,acetonitrile, and isopropanol conversion to Form D was observed in allsolvents. Without wishing to be bound by theory, these results as wellas conversion of Form F to Form D in these conditions suggest that FormD is more thermodynamically stable than Form C and F according toOstwald's Rule of Stages. A thermal-stability experiment on Form C at60° C. found the form to be stable. Conversion to another form was notobserved.

6.2.11.5. Characterization of Polymorphic Form D of Diazoxide PotassiumSalt.

Form D of diazoxide potassium salt is a hemihydrate. It is a crystallineform of diazoxide potassium, with endothermic events at 130, 191, and352° C. in the DSC. FTIR (ATR) spectroscopy showed differences betweenforms. ¹H NMR spectra were found to be consistent with the structure ofdiazoxide. NMR data also indicated that the magnetic environment of thediazoxide structure had changed evidenced by a movement in chemicalshift of the aromatic and methyl proton resonances. In addition, theresonance due to the amine proton was not observed which suggesteddeprotonation in solution. Weight loss by TGA was 4.5%, consistent witha half-molar equivalent of water, and occurred near 110° C. consistentwith the endotherm observed in the DSC experiment. Moisture-sorptionanalysis conducted at 25° C. from 0 to 90% RH (adsorption) and 85 to 0%RH (desorption) showed Form D to be a hygroscopic, showing approximately26 wt % water at 90% RH. In comparison, Forms C and F (acetone anddioxane solvates) showed approximately 47 and 22 wt % water at 90% RH,respectively. XRPD analysis following the moisture-sorption experimentafforded a pattern consistent with Form D. Solubility studies wereconducted at pH 2, 7, and 12 for Form D and showed 29, 33, and 59 mg/mLrespectively. Solubility concentrations were determined usingarea-percent calculations with an HPLC calibration curve. Slurryexperiments were performed on 50/50 mixtures of Forms C, D, and F todetermine their propensity for conversion and to see if a unique formcould be generated. Upon slurrying mixtures of Form D with Form C orForm F in ethyl acetate, acetonitrile, and isopropanol conversion toForm D was observed in all solvents.

6.2.11.6. Characterization of Polymorphic Form F of Diazoxide PotassiumSalt.

Form F of diazoxide potassium salt is a dioxane solvate with a 2:1 ratioof diazoxide/solvent. It is a crystalline form of diazoxide potassium,with endothermic events at 191 and 363° C. in the DSC. The XRPD patternfor Form F is unique compared to all other forms observed. FTIR (ATR)spectroscopy showed differences between forms. ¹H NMR spectra were foundto be consistent with the structure of diazoxide with a half-molarequivalent of dioxane present. NMR data also indicated that the magneticenvironment of the diazoxide structure had changed as evidenced by amovement in chemical shift of the aromatic and methyl proton resonances.In addition, the resonance due to the amine proton was not observedwhich suggested deprotonation in solution. Weight loss by TGA was 13.1%,consistent with a half-molar equivalent of dioxane, and occurs near 180°C. consistent with the endotherm observed in the DSC experiment.Moisture-sorption analysis conducted at 25° C. from 0 to 90% RH(adsorption) and 85 to 0% RH (desorption) showed Form F to be ahygroscopic solid, showing approximately 22 wt % water at 90% RH. Incomparison, Forms C and D (acetone solvate and hemihydrate) showedapproximately 47 and 26 wt % water at 90% RH, respectively. XRPDanalysis following the moisture-sorption experiment afforded a patternconsistent with Form D. Slurry experiments were performed on 50/50mixtures of Forms C, D, and F to determine their propensity forconversion and to see if a unique form could be generated. Uponslurrying mixtures of Form F with Form C and Form D in ethyl acetate,acetonitrile, and isopropanol conversion to Form D was observed in allsolvents.

B. In Vivo Obesity Testing

1. Obesity Animal Model

Formulations of salts of any of the compounds of Formulae I-IV preparedas described herein can be tested for efficacy in an animal model ofobesity as described by Surwit et al. (Endocrinology 141:3630-3637(2000)). Briefly, 4-week-old B6 male mice are housed 5/cage in atemperature-controlled (22° C.) room with a 12-h light, 12-h dark cycle.The high fat (HF) and low fat (LF) experimental diets contain 58% and11% of calories from fat, respectively. A group of mice are fed the HFdiet for the first 4 weeks of the study; the remaining 15 mice are fedthe LF diet. The mice assigned to the LF diet are maintained on thisdiet throughout the study as a reference group of lean control mice. Atweek 4, all HF-fed mice a reassigned to 2 groups of mice. The firstgroup remains on the HF diet throughout the study as the obese controlgroup. The remaining 3 groups of mice are fed the HF diet andadministered the controlled release formulation of salts of any of thecompounds of Formulae I-IV at about 150 mg of active per kg per day as asingle dose administered by oral gavage. Animals are weighed weekly, andfood consumption is measured per cage twice weekly until the diets arechanged at week 4, whereupon body weight and food intake are determineddaily. The feed efficiency (grams of body weight gained per Calconsumed) is calculated on a per cage basis. Samples for analysis ofinsulin, glucose, and leptin are collected on day 24 (4 days before thediets are changed), on day 32 (4 days after the change), and biweeklythereafter. In all cases food is removed 8 h before samples arecollected. Glucose is analyzed by the glucose oxidase method. Insulinand leptin concentrations are determined by double antibody RIA. Theinsulin assay is based on a rat standard, and the leptin assay uses amouse standard. At the termination of the study, a postprandial plasmasample is collected and analyzed for triglyceride and nonesterifiedfatty acid concentrations. After 4 weeks of drug treatment, a subset of10 animals from each group is killed. The epididymal white adiposetissue (EWAT), retroperitoneal (RP) fat, interscapular brown adiposetissue (IBAT) fat pads, and gastrocnemius muscle are removed, trimmed,and weighed. The percent body fat is estimated from the weight of theepididymal fat pad. A subset of five animals from each group is injectedi.p. with 0.5 g/kg glucose. At 30 min post injection, a plasma sample iscollected and analyzed for glucose content by the glucose oxidasemethod.

2. Treatment of Obesity in Humans

Formulations of salts of any of the compounds of Formulae I-IV preparedas described herein can be tested for efficacy in obese humans, asdescribed by Alemzadeh (Alemzadeh et al., J Clin Endocr Metab83:1911-1915 (1998)). Subjects consist of moderate-to-morbidly obeseadults with a body mass index (BMI) greater than or equal to 30 kg/m².Each subject undergoes a complete physical examination at the initialevaluation, body weight being measured on a standard electronic scaleand body composition being measured by DEXA.

Before initiation of the study, all subjects are placed on a hypocaloricdiet for a lead-in period of 1 week. This is designed to excludesubjects who are unlikely to be compliant and to ensure stable bodyweight before treatment. Up to 50 subjects are tested at each dosage ofdrug. Daily dosage is set at 100, 200, and 300 mg/day. The daily dose isdivided into 2 doses for administration. The dose is administered aseither one, two or three 50 mg capsules or tablets at each time ofadministration. Subjects are dosed daily for up to 12 months. Subjectsare reviewed weekly, weighed, and asked about any side effects orconcurrent illnesses.

Twenty-four-hour dietary recall is obtained from each subject. Thedietary recalls are analyzed using a standard computer software program.All subjects are placed on a hypocaloric diet and encouraged toparticipate in regular exercise.

Before commencing, and after completion of the study, the followinglaboratory tests are obtained: blood pressure, fasting plasma glucose,insulin, cholesterol, triglycerides, free fatty acids (FFA), andglycohemoglobin, and measures of rate of appearance and oxidation ofplasma derived fatty acids. Additionally, routine chemistry profiles andfasting plasma glucose are obtained weekly to identify those subjectswith evidence of glucose intolerance and/or electrolyte abnormalities.Glucose is analyzed in plasma, by the glucose oxidase method.

Insulin concentration is determined by RIA using a double-antibody kit.Cholesterol and triglycerides concentrations are measured by anenzymatic method. Plasma FFA is determined by an enzymatic colorimetricmethod. SI was assessed by an iv glucose tolerance test (IVGTT) usingthe modified minimal model. After an overnight fast, a glucose bolus(300 mg/kg) was administered iv, followed (20 min later) by a bolus ofinsulin. Blood for determination of glucose and insulin is obtained froma contra lateral vein at −30, −15, 0, 2, 3, 4, 5, 6, 8, 10, 19, 22, 25,30, 40, 50, 70, 100, 140, and 180 min. SI and glucose effectiveness (SG)are calculated using Bergman's modified minimal-model computer programbefore and after the completion of the study. Acute insulin response toglucose is determined over the first 19 min of the IVGTT, and theglucose disappearance rate (Kg) is determined from 8-19 min of theIVGTT. Body composition is measured by bioelectrical impedance beforeand at the completion of the study. Resting energy expenditure (REE) ismeasured by indirect calorimetry after an overnight 12-h fast, withsubjects lying supine for a period of 30 min. Urine is collected overthe corresponding 24 h, for measurement of total nitrogen anddetermination of substrate use, before and after the study.

3. Treatment of Obesity in Humans by Coadministering Diazoxide andPhentermine

Evaluation of a prolonged co-administration of solid oral dosage form ofsalts of any of the compounds of Formulae I-IV thereof in combinationwith phentermine can be conducted in humans with moderate-to-morbidobesity and a body mass index (BMI) greater than or equal to 30 kg/m².Each subject undergoes a complete physical examination at the initialevaluation, body weight being measured on a standard electronic scaleand body composition by DEXA.

Before initiation of the study, all subjects are placed on a hypocaloricdiet for a lead-in period of 1 week. This is designed to excludesubjects who are unlikely to be compliant and to ensure stable bodyweight before treatment. Up to 100 subjects are tested. Daily dosage ofsalts of any of the compounds of Formulae I-IV is set at 200 mg. Thedaily dose is divided into 2 doses for administration. The dose isadministered as either a 100 mg capsule or a 100 mg tablet at each timeof administration. Subjects are dosed daily for up to 12 months.Phentermine is administered as a single daily dose of 15 mg. Subjectsare reviewed every two weeks, weighed, and asked about any side effectsor concurrent illnesses.

All subjects are continued on a hypocaloric diet and encouraged toparticipate in regular exercise. Before commencing, and after completionof the study, laboratory tests as described in the example above areobtained.

4. Prevention of Diabetes in Prediabetic Humans

The example describes use of salts of any of the compounds of FormulaeI-IV in a prediabetic subject to prevent the occurrence of diabetes.Subjects included in the study all have elevated risk of developingdiabetes as measured by one of two methods. In a fasting glucose assaythey have plasma glucose values between 100 and 125 mg/dl indicatingimpaired fasting glucose, or in an oral glucose tolerance test they haveplasma glucose values between 140 and 199 mg/dl at 2 hours post-glucoseload indicating they have impaired glucose tolerance. Treatment isinitiated in any subject meeting either criteria. Treated subjectsreceive either 200 mg diazoxide per day as a 100 mg capsule or tablettwice per day or as two 100 mg capsules or tablets once per day. Placebotreated subjects receive either one placebo capsule or tablet twice perday or two placebo capsules or tablets once per day.

Treatment is continued for once year with OGTT or fasting glucosemeasured monthly.

5. A Sustained Release Coformulation of Diazoxide HCl and Metformin HClUse to Treat Diabetic Patients

A sustained release co-formulation of diazoxide HCl and metformin HCl isproduced by forming a compressed tablet matrix that includes 750 mg ofmetformin HCl and 100 mg of diazoxide HCl. These active ingredients areblended with sodium carboxymethyl cellulose (about 5% (w/w)),hypromellose (about 25% (w/w), and magnesium stearate (<2% (w/w)). Thecompressed tablet is further coated with a combination of ethylcellulose(80% (w/w)) and methyl cellulose (20% (w/w)) as a thin film to controlrate of hydration and drug release.

Type II diabetic patients are treated with the oral dosage form byadministration of two tablets once per day or one tablet every 12 hours.Treatment of the patient with the drug is continued until one of twotherapeutic endpoints is reached, or for so long as the patient derivestherapeutic benefit from administration. The two therapeutic endpointsthat would serve as the basis for the decision to cease treatmentinclude the patient reaching a Body Mass Index (BMI (kg/m²)) between 18and 25 or the re-establishment of normal glucose tolerance in theabsence of treatment. The patient is monitored periodically for (a)glucose tolerance using an oral glucose tolerance test, (b) glycemiccontrol using a standard blood glucose assay, (c) weight gain or loss,(d) progression of diabetic complications, and (e) adverse effectsassociated with the use of these active ingredients.

6. Prevention or Treatment of Weight Gain in a Patient Treated withOlanzapine

Pharmacotherapy for schizophrenia is initiated for a patient meeting DSMIII-R criteria for schizophrenia. The patient is administered 10 mg ofolanzapine (Zyprexa, Lilly) once per day. Adjunctive therapy to thepatient for schizophrenia includes 250 mg equivalent of valproic acid asdivalproex sodium (Depakote, Abbott Labs). Weight gain, dyslipidemia andimpaired glucose tolerance, and metabolic syndrome are high frequencyadverse events in patients treated with this combination ofanti-psychotics. Weight gain, dyslipidemia, impaired glucose toleranceor metabolic syndrome are treated by the co-administration of atherapeutically effective dose of a K_(ATP) channel opener. The patientis treated with administration of 200 mg/day of salts of any of thecompounds of Formulae I-IV as a once daily tablet formulation.Administration of salts of any of the compounds of Formulae I-IVcontinues until the weight gain, dyslipidemia, impaired glucosetolerance or metabolic syndrome is corrected or until treatment of thepatient with olanzapine is discontinued. Dyslipidemia is detected bymeasuring circulating concentrations of total, HDL, and LDL cholesterol,triglycerides and non-esterified fatty acids. Impaired glucose toleranceis detected through the use of oral or IV glucose tolerance tests.Metabolic syndrome is detected by measuring its key risk factorsincluding central obesity, dyslipidemia, impaired glucose tolerance, andcirculating concentrations of key proinflammatory cytokines

7. Comparison of Single Doses of Diazoxide Administered as Proglycem®Oral Suspension or as Diazoxide Choline Controlled-Release Tablets inObese Subjects.

7.1 Experimental Design

7.1.1. Objective of Study.

Clinical studies employing a randomized, open-label, parallel protocolcomparing the safety, tolerability and bioavailability (i.e.,pharmacokinetics) in obese subjects of Proglycem® (oral suspension) anddiazoxide choline salt (controlled-release tablet) were conducted. Thestudy evaluated the safety and tolerability of a single 200 mg does(approximately 2 mg/kg) of diazoxide. The study further compared thesingle dose pharmacokinetics of an oral suspension of the free base ofdiazoxide (Proglycem®) with a controlled-release tableted formulation ofdiazoxide choline salt under fasting conditions in obese subjects.

7.1.2. Rationale for Study.

Diazoxide choline was selected as an alternative molecule for the oraladministration of diazoxide because of significantly greater aqueoussolubility over diazoxide free base, rapid conversion to diazoxide baseon exposure to an aqueous environment, and incompatibility withincorporation into sustained release formulations. The fate of diazoxidecholine in an aqueous medium in vitro has been extensivelycharacterized. Without wishing to be bound by theory, once solubilizedfrom the controlled-release tablet formulation, prior to absorption, thesalt hydrolyzes to the free base of diazoxide and choline hydroxide.This was extensively characterized using UV/Vis absorbance, and occurredat all physiological relevant pH values from pH 2.0 to pH 8.5. Thishydrolysis occurred in deionized water and buffered aqueous solutions,and in polar solvents that contained trace amounts of water. Thus,diazoxide, as the free base, is the molecular form absorbed followingoral administration of the choline salt of diazoxide to animals orhumans. Serum and plasma assays used in TK and PK analysis measurediazoxide free base concentrations. Dissolution assays measure diazoxidefree base. Differences in plasma concentration-time profiles ofdiazoxide will be dependent on the time course of release of diazoxidecholine from the tablet matrix in the intestinal tract. Historically,both oral suspension and immediate release capsule formulations ofdiazoxide have been commercially available, marketed as Proglycem®. Theoral suspension has been shown to be highly bioavailable and rapidlyabsorbed upon administration. Because of the difficulty incharacterizing dissolution from the oral suspension, the in-vitrodissolution of Proglycem® capsules was compared under standardizedconditions to that of diazoxide choline controlled-release tablets (50mg and 200 mg).

7.1.3. Inclusion Criteria.

Inclusion criteria for the present study included age 18 to 65 years oldinclusive and BMI between 30 and 45 kg/m², inclusive, and signedinformed consent. Female participants were required to be eitherpostmenopausal for at least 1 year, surgically sterile [bilateral tuballigation, bilateral oophorectomy, or hysterectomy], or practicing amedically acceptable method of birth control. Inclusion criteria furtherincluded freedom from serious medical disorders involving the kidneys,digestive system, heart and blood vessels, lungs, liver, eyes, nerves,brain, skin, endocrine system, bones or blood. Subjects, other thantheir obese condition, were generally healthy as documented by medicalhistory, physical examination, vital sign, 12-lead ECG, and clinicallaboratory assessments. Characteristics of the subjects randomized inthe study are provided in Table 33.

TABLE 33 Characteristics of subjects randomized in the study ProglycemOral Diazoxide Choline Controlled- Suspension (n = 15) Release Tablet (n= 15) Parameter Mean ± SD (range) Mean ± SD (range) Age (yr) 32.5 ± 12.1(18-56) 27.9 ± 12.6 (19-54) BMI 32.9 ± 4.3 (30.2-44.8) 33.8 ± 3.2(31.4-42.1) (kg/m²) Gender 9/6 7/8 (male/ female)

7.1.4. Exclusion Criteria.

Exclusion criteria included treatment with an investigational drugwithin 28 days prior to dosing, presence or history of a clinicallysignificant disorder, clinical laboratory test values outside of theaccepted reference range, reactive screen for HBV, HCV, or HIV, use ofany medication affecting body weight, lipid or glucose metabolism within2 months, use of any systemic prescription medication from 14 days priorto screening to dosing except hormonal contraceptives, use of any drugknown to induce or inhibit hepatic drug metabolism from 28 days prior toscreening until dosing, positive test for drug of abuse, current tobaccouse, positive pregnancy screen, or pregnant or breastfeeding.

7.1.5. Randomization.

A total of 30 subjects were randomized in the present study. Fifteensubjects were randomized to each arm. For convenience, the 30 subjectswere broken up into two cohorts. Cohort 1 checked into the clinic onOct. 12, 2006, was dosed on Oct. 14, 2006, remained in the clinic for 72hours following dose administration, returning at 96 and 120 hours afterdose administration. Cohort 1 included 10 subjects randomized to receiveProglycem Oral Suspension and 5 subjects randomized to receive aDiazoxide Choline Controlled-Release Tablet. Cohort 2 checked into theclinic on Oct. 14, 2006, were dosed on Oct. 16, 2006, remained in theclinic for 72 hours following dose administration, returning at 96 and120 hours after dose administration. Cohort 2 included 5 subjectsrandomized to receive Proglycem® Oral Suspension and 10 subjectsrandomized to receive a diazoxide choline controlled-release tablet. Allsubjects completed the study.

7.1.6. Dosing.

Subjects randomized to the Proglycem® Oral Suspension arm received asingle 200 mg dose (4 mL) taken with 240 mL of room temperature water.Subjects randomized to the diazoxide choline controlled-release tabletarm received a single tablet containing 290 mg of diazoxide choline,which is equivalent to 200 mg of diazoxide. Doses were administeredafter an overnight fast, and fasting continued until approximately 4.25hours after dose administration at which time a standardized meal wasserved.

7.1.7. Safety Monitoring.

Clinical laboratory tests including hematology, clinical serum chemistryand urinalysis were conducted at screening and at end of study.Hematology included measurements of hemoglobin, hematocrit, white bloodcell count with differential, red blood cell count and platelet count.Clinical serum chemistry included evaluation of sodium, potassium, BUN,creatinine, total bilirubin, total protein, albumin, alkalinephosphatase, AST, ALT, glucose, total cholesterol, HDL-cholesterol, andtriglycerides. Urinalysis included pH, specific gravity, protein,glucose, ketones, bilirubin, blood, nitrites, urobilinogen, leukocytes,and microscopic urine analysis if sample was dipstick positive. Alladverse events were recorded. Vital signs were collected at screening,at one hour prior to dosing, and after dose administration at 1, 3, 6,9, 12, 24, 48, 72 and 120 hours. Continuous two-lead cardiac monitoring(telemetry) was performed on subjects from 24 hours prior to doseadministration (to establish baseline data) until 24 hours after doseadministration.

7.1.8. Exploratory Endpoints.

Three exploratory endpoints were evaluated in the study. Blood glucose,insulin and non-esterified fatty acids (NEFA) were measured prior todosing and at 1, 3, 6, 9, 12, 24, and 48 hours post dosing. Bloodglucose and insulin levels were also assessed at end of study. Datacollected at times 1, 3, 24, and 48 hours after dose administration wereobtained under fasting conditions. The remaining data points werepost-prandial measures. Samples for pharmacokinetic analysis to evaluatebioavailability were collected prior to dosing and after doseadministration at 1, 2, 4, 6, 8, 12, 16, 20, 24, 32, 40, 48, 72, 96, and120 hours.

7.2. Results

7.2.1. Adverse Events.

The adverse events observed with each of the two formulations aresummarized in Tables 34 and 35. With the single exception, a moderateheadache requiring treatment with acetaminophen in a subject treatedwith Proglycem® Oral Suspension, all adverse events were mild. Bothformulations appeared to be well tolerated in the study. One subjectadministered with diazoxide choline controlled-release tabletexperienced mild loss of appetite. Because most obese and obese diabeticanimal model studies of diazoxide show some reduction in feedconsumption, loss of appetite was, therefore, considered part of thepharmacodynamic response to the drug in obese patients, rather than anadverse event.

TABLE 34 Adverse events (AE) in subjects administered Proglycem ® OralSuspension (n = 15) Hours Post Corrective Out- Adverse Event DosingTreatment come Gastrointestinal Nausea^(a) 2.75 Mild None ResolvedNeurological Headache 20.0 Moderate Therapy required Resolvedacetaminophen Headache^(b) 6.25 Mild None Resolved Dizziness^(c) 1.0Mild Assisted to supine Resolved position Dizziness^(c) 3.25 MildAssisted to supine Resolved position Dizziness^(d) 1.0 Mild Assisted tosupine Resolved position Dizziness^(d) 4.25 Mild Assisted to supineResolved position Miscellaneous Chills^(b) 6.25 Mild None ResolvedDermatitis from 4.75 Mild None Resolved ECG patches^(a) Number ofsubjects with at least one AE = 5, AEs followed by the same superscriptletter are the same subject

TABLE 35 Adverse events (AE) in subjects administered Diazoxide CholineControlled-Release Tablet (n = 15) Hours Post Corrective Out- AdverseEvent Dosing Treatment come Gastrointestinal Nausea^(e) 1.5 Mild NoneResolved Dry Heaves^(e) 1.6 Mild None Resolved Neurological Lightheaded12.0 Mild Assisted to supine Resolved position Dizziness^(g) 20.0 MildNone Resolved Miscellaneous Chills^(f) 2.0 Mild None Resolved BackPain^(f) 2.0 Mild None Resolved Warm^(g) 20.0 Mild None Resolved Numberof subjects with at least one AE = 4, AEs followed by the samesuperscript letter are the same subject

7.2.2. Clinical Chemistry.

A summary of fasting glucose, fasting insulin, NEFA, and serum sodium(Na), potassium (K) and creatinine level is provided in Table 36.Neither treatment resulted in significant changes in serum Na, K orcreatinine from baseline to end of study. Treatment with diazoxideprimarily impacts glucose-stimulated insulin secretion rather than basalinsulin secretion. Fasting glucose levels increased slightly in thefirst 3 hours following dose administration. This increase was morepronounced in the Proglycem® Oral Suspension arm as compared to thediazoxide choline controlled-release tablet arm. These results areconsistent with the differences in measured rates of dissolution fromthese formulations (see Table 5 herein) and the PK levels (see FIGS. 24and 25 herein). In this study, there is no evidence for a reduction infasting insulin after the administration of the single dose ofdiazoxide. There is also no evidence for a significant increase infasting glucose as measured at 24 or 48 hours after dose administration.The most sensitive pharmacodynamic response measure is NEFA. Diazoxidetreatment normally results in a short-term increase in NEFA, whichreturns to levels at or below baseline in about 6 hours. Bothformulations showed this transient increase in NEFA. Treatment withProglycem® Oral Suspension resulted in a statistically significantincrease (p<0.001) in NEFA at 3 hours, which by 6 hours after doseadministration had returned to levels well below baseline. Similarly theincrease in NEFA at 3 hours after administration of diazoxide cholinecontrolled-release tablets was statistically significant (p<0.0012).NEFA levels in the diazoxide choline controlled-release tablet treatedsubjects also returned to levels well below baseline by 6 hours afterdose administration.

TABLE 36 Fasting glucose, fasting insulin, non-esterified fatty acid(NEFA), serum sodium (Na), potassium (K) and creatinine of subjectsDiazoxide Choline Proglycem Oral Controlled-Release Suspension (n = 15)Tablet (n = 15) Parameter Mean ± SD (range) Mean ± SD (range) FastingGlucose (mg/dL) Baseline 93 ± 6 (83-104) 93 ± 6 (83-108) 1 h post-dose99.4 ± 7 (89-117) 97.9 ± 7 (92-117) 3 h post-dose 102.8 ± 6 (93-113)98.0 ± 7 (90-112) 24 h post-dose 97 ± 5 (87-106) 98 ± 8 (88-118) 48 hpost-dose 92 ± 5 (84-103) 93 ± 5 (83-104) Fasting Insulin (μIU/ml)Baseline 8.6 ± 5.2 (2.9-24.1) 9.9 ± 3.3 (4.4-16.3) 24 h post-dose 12.0 ±8.9 (6.1-41.9) 11.1 ± 3.3 (5.5-17.0) 48 h post-dose 9.5 ± 5.7 (4.0-28.9)11.1 ± 2.9 (5.3-17.8) NEFA (μmol/L) 1 h post-dose 0.36 ± 0.11(0.21-0.65) 0.34 ± 0.14 (0.09-0.67) 3 h post-dose 0.53 ± 0.13(0.31-0.78) 0.55 ± 0.19 (0.28-0.88) 6 h post-dose 0.13 ± 0.07(0.07-0.32) 0.11 ± 0.04 (0.07-0.21) 24 h post-dose 0.36 ± 0.10(0.23-0.62) 0.40 ± 0.18 (0.25-0.97) 48 h post-dose 0.38 ± 0.13(0.2-0.66) 0.39 ± 0.15 (0.17-0.65) Na (mEq/L) Baseline 138.0 ± 2.0(135-142) 138.1 ± 2.6 (134-143) End of Study 138.7 ± 1.1 (137-141) 139.3± 2.3 (135-144) K (mmol/L) Baseline 4.4 ± 0.4 (3.9-5.5) 4.2 ± 0.3(3.8-5.1) End of Study 4.4 ± 0.3 (3.9-5.2) 4.2 ± 0.3 (3.8-5.2)Creatinine (mg/dL) Baseline 0.9 ± 0.1 (0.7-1.1) 0.9 ± 0.2 (0.7-1.4) Endof Study 1.0 ± 0.1 (0.8-1.3) 0.9 ± 0.2 (0.6-1.3)

All other clinical laboratory tests, including hematology, clinicalserum chemistry, and urinalysis were within normal ranges for obesesubjects.

7.2.3. Vital Signs.

A graph depicting sitting blood pressure at baseline and at varioustimes following dose administration is provided in FIG. 22. Neithertreatment was associated with a sustained reduction or increase in bloodpressure, nor was there any clear trend from baseline over 6, 9, 12, or24 hours after dose administration. Pulse rates dropped incrementallyfrom baseline levels in the period from dose administration to 3 hourspost dosing (FIG. 23). Pulse rates rose to levels equivalent to baselineor slightly above baseline in the period from 6 to 12 hours after doseadministration, and remained at baseline levels from 24 hours after doseadministration to the end of the study (FIG. 23). Review of the baselinecardiac telemetry data for all subjects was conducted the morning ofdosing. Based on the absence of clinical significant abnormalities, allsubjects were allowed to proceed with dosing. The cardiac telemetry datafrom study time 0 to 24 hours after dose administration was evaluated.No clinically significant abnormalities (e.g. arrhythmias) wereobserved.

7.2.4. Preliminary Evaluation of Diazoxide Plasma Pharmacokinetics.

Compared to the Proglycem® Oral Suspension, the diazoxide cholinecontrolled-release tablet had a 30% lower peak exposure (Cmax) and a 15%lower total exposure (AUC) when the formulations were administeredorally as 200 mg equivalents of diazoxide (Table 37). The timing of thepeak plasma concentration (Tmax) occurred at a median time of 4 hr afteradministration of the oral suspension and 20 hr after administration ofthe diazoxide choline controlled-release tablet. The terminal half-lifefor diazoxide was similar with both formulations (29 hr for the oralsuspension and 32 hr for the tablet). The two formulations had similarbetween-patient variability for Cmax andAUC. The most notable differencebetween the two formulations following a single dose was the lower,broader peak of the concentration-time profile of the tablet formulation(FIGS. 24 and 25). The peak concentration for the tablet averaged about30% less and occurred about 16 hours later than the suspension. FIGS. 24and 25 illustrate that the mean peak concentration was virtuallyunchanged between 12-hr and 24-hr post-dose following administration ofthe diazoxide choline controlled-release tablet formulation. Because ofa more sustained release pattern, the tablet formulation was predictedto have a greater accumulation factor with chronic dosing than was theoral suspensions (3.96 vs. 2.84). Simulations of repeated once-dailydosing with the two formulations (assuming linear pharmacokinetics)predicted that the two formulations would have similar troughconcentrations at steady-state (22-23 μg/mL) (see FIG. 26).

TABLE 37 Summary Statistics for Plasma Diazoxide PharmacokineticParameters after a Single Dose of Two Formulations Cmax Tmax AUC(0-24)AUC(0-120) AUC(0-∞) λz T½ Accum μg/mL hr μg · hr/mL μg · hr/mL μg ·hr/mL 1/hr hr Factor Diazoxide Choline Controlled-Release Tablets N 1515 15 15 15 15 15 15 Mean 9.25 22.1 166 553 618 0.0242 31.9 3.96 Geomean9.07 19.6 157 531 588 0.0229 30.2 3.74 St Dev 1.89 11.5 54 160 1980.0085 10.6 1.48 SEM 0.49 3.0 14 41 51 0.0022 2.7 0.38 CV % 20.5% 52.0%32.5% 28.9% 32.0% 35.2% 33.3% 37.5% Median 9.08 20.0 164 518 586 0.023729.3 3.80 Min 5.53 8.00 74.6 261 274 0.0144 15.9 2.26 Max 13.2 48.0 266842 1024 0.0437 48.3 7.84 Diazoxide Oral Suspension (Proglycem) N 15 1515 15 15 15 15 15 Mean 13.5 6.67 245 643 696 0.0256 28.6 2.84 Geomean13.3 5.36 243 629 677 0.0249 27.9 2.79 St Dev 2.3 5.49 35 142 173 0.00686.3 0.57 SEM 0.6 1.42 9 37 45 0.0018 1.6 0.15 CV % 17.1% 82.3% 14.4%22.1% 24.9% 26.7% 22.0% 20.0% Median 13.3 4.00 245 620 671 0.0234 29.62.86 Min 10.1 2.00 183 464 488 0.0180 16.1 1.84 Max 18.0 24.0 314 9261054 0.0430 38.5 4.25

7.3 Conclusions of Comparison of Single Doses of Diazoxide Administeredas Proglycem® Oral Suspension or as Diazoxide Choline Controlled-ReleaseTablets in Obese Subjects.

The present clinical study on the comparison of a single dose ofdiazoxide choline controlled-release tablet with an equivalent dose ofdiazoxide administered as Proglycem® Oral Suspension in obese patientsindicates that both formulations were well tolerated. With a singleexception, moderate headache requiring treatment with acetaminophen inone subject treated with Proglycem®, all adverse events were mild.Diazoxide choline controlled-release tablets appear to have a better CNSsafety profile than Proglycem® Oral Suspension as evidenced by theabsence of headaches and reduced rate of dizziness. Exploratoryendpoints showed no detrimental impact. Preliminary pharmacokineticanalysis showed that compared to Proglycem® Oral Suspension, thediazoxide choline controlled-release tablet had a 30% lower peakexposure (Cmax) and a 15% lower total exposure (AUC) for the same 200 mgdiazoxide equivalent dose. The timing of the peak plasma concentration(Tmax) occurred at a median time of 4 hr after administration of theoral suspension and 20 hr after administration of the diazoxide cholinetablet. The terminal half-life for diazoxide is similar with bothformulations (29 hr for the oral suspension and 32 hr for the tablet).The two formulations had similar between-patient variability for CmaxandAUC.

All patents and other references cited in the specification areindicative of the level of skill of those skilled in the art to whichthe invention pertains, and are incorporated by reference in theirentireties, including any tables and figures, to the same extent as ifeach reference had been incorporated by reference in its entiretyindividually.

One skilled in the art would readily appreciate that the presentinvention is well adapted to obtain the ends and advantages mentioned,as well as those inherent therein. The methods, variances, andcompositions described herein as presently representative of preferredembodiments are exemplary and are not intended as limitations on thescope. Changes therein and other uses will occur to those skilled in theart, which are encompassed within the spirit of the invention, aredefined by the scope of the claims.

Definitions provided herein are not intended to be limiting from themeaning commonly understood by one of skill in the art unless indicatedotherwise.

The inventions illustratively described herein may suitably be practicedin the absence of any element or elements, limitation or limitations,not specifically disclosed herein. Thus, for example, the terms“comprising”, “including,” containing”, etc. shall be read expansivelyand without limitation. Additionally, the terms and expressions employedherein have been used as terms of description and not of limitation, andthere is no intention in the use of such terms and expressions ofexcluding any equivalents of the features shown and described orportions thereof, but it is recognized that various modifications arepossible within the scope of the invention claimed. Thus, it should beunderstood that although the present invention has been specificallydisclosed by preferred embodiments and optional features, modificationand variation of the inventions embodied therein herein disclosed may beresorted to by those skilled in the art, and that such modifications andvariations are considered to be within the scope of this invention.

The invention has been described broadly and generically herein. Each ofthe narrower species and subgeneric groupings falling within the genericdisclosure also form part of the invention. This includes the genericdescription of the invention with a proviso or negative limitationremoving any subject matter from the genus, regardless of whether or notthe excised material is specifically recited herein. Other embodimentsare within the following claims. In addition, where features or aspectsof the invention are described in terms of Markush groups, those skilledin the art will recognize that the invention is also thereby describedin terms of any individual member or subgroup of members of the Markushgroup.

That which is claimed is:
 1. A pharmaceutical formulation comprising: i.) a salt of a K_(ATP) channel opener selected from the group consisting of Formula III and Formula IV, and a counter ion as shown below:

wherein: R¹ is selected from the group consisting of hydrogen, lower alkyl, substituted lower alkyl, and cycloalkyl provided however that when R¹ is a substituted lower alkyl, then the substituent does not include an amino group; R^(2a) is hydrogen; R^(2b) is hydrogen; R³ is selected from the group consisting of hydrogen, halogen, lower alkyl, substituted lower alkyl, cycloalkyl and substituted cycloalkyl provided however that when R³ is a substituted lower alkyl, then the substituent does not include an amino group; R⁴ is selected from the group consisting of hydrogen, halogen, lower alkyl, substituted lower alkyl, cycloalkyl and substituted cycloalkyl provided however that when R⁴ is a substituted lower alkyl, then the substituent does not include an amino group, and ii.) a cholesterol lowering drug.
 2. The formulation of claim 1 wherein said K_(ATP) channel opener is diazoxide.
 3. The formulation of claim 1 wherein said K_(ATP) channel opener is diazoxide and said counter ion is choline.
 4. The formulation of claim 1 wherein said K_(ATP) channel opener is diazoxide and said counter ion is hexamethyl hexamethylene diammonium.
 5. The formulation of claim 1 wherein said formulation is a sustained release formulation.
 6. The formulation of claim 5 wherein said sustained release formulation has a period of release 8 to 24 hours following administration.
 7. The formulation of claim 5 wherein said sustained release formulation has a period of release 2 to 30 hours following administration.
 8. The formulation of claim 1, wherein said cholesterol lowering drug is a statin.
 9. The formulation of claim 8, wherein said statin is selected from the group consisting of pravastin, simvastatin, fluvastatin, rosuvastin, and lovastatin.
 10. The formulation of claim 8, wherein said pharmaceutical formulation is formulated for once a day administration, and said formulation comprises about 10 to 80 mg/unit of said statin.
 11. The formulation of claim 10, wherein said formulation comprises about 50 to 400 mg/unit of said salt of a K_(ATP) channel opener.
 12. The formulation of claim 8, wherein said pharmaceutical formulation is formulated for twice a day administration, and said formulation comprises about 5 to 40 mg/unit of said statin.
 13. The formulation of claim 12, wherein said formulation comprises about 25 to 200 mg/unit of the salt of a K_(ATP) channel opener.
 14. The formulation of claim 1 for a single administration that contains between 10 and 2000 mg of the salt.
 15. The formulation of claim 1 for a single administration that contains between 200 and 500 mg of the salt.
 16. The formulation of claim 1 wherein said formulation is a delayed release formulation.
 17. The formulation of claim 16 wherein said delayed release formulation further comprises at least one component that substantially inhibits release of the K_(ATP) channel opener until after gastric transit.
 18. The formulation of claim 16 wherein said delayed release formulation further comprises a component selected from the group consisting of: (a) a pH sensitive polymer or co-polymer applied as a compression coating on a tablet; (b) a pH sensitive polymer or co-polymer applied as a thin film on a tablet; (c) a pH sensitive polymer or co-polymer applied as a thin film to an encapsulation system; (d) a pH sensitive polymer or co-polymer applied to encapsulated microparticles; (e) a non-aqueous-soluble polymer or copolymer applied as a compression coating on a tablet; (f) a non-aqueous-soluble polymer or co-polymer applied as a thin film on a tablet; (g) a non-aqueous soluble polymer applied as a thin film to an encapsulation system, and (h) a non-aqueous soluble polymer applied to microparticles; wherein the pH sensitive polymer or co-polymer of (a), (b), (c), and (d) is resistant to degradation under acid conditions.
 19. The formulation of claim 16 wherein said delayed release formulation exhibits sustained release of the K_(ATP) channel opener following gastric transit.
 20. The formulation of claim 19 wherein the formulation comprises a component selected from the group consisting of: (a) a pH sensitive polymeric coating; (b) a hydrogel; (c) a film coating that controls the rate of diffusion of the salt from a coated matrix; (d) an erodable matrix that controls rate of salt release; (e) polymer coated pellets, granules or microparticles of salt which can be further encapsulated or compressed into a tablet; (f) an osmotic pump system containing the salt; (g) a compression coated tablet form of the salt; and (h) combinations thereof. 